The VirB System Plays a Crucial Role in Brucella Intracellular Infection
Round 1
Reviewer 1 Report
Title:
Dear editor:
To explore the mechanisms underlying the infection of Brucella spp. on target organisms is a very interesting topic. In the manuscript submitted by Xiong et al., the authors try to explore and reveal the mechanisms underlying this process. However, in many of the paragraphs there is still a discussion made by the authors in order to summarize or draw a final idea from the delivered information. More citations for figures must be added in the manuscript body, and the resolution of the figures must be enhanced. In addition, I think there are a large number of abbreviations (some of them unnecessary) that make the reading of the manuscript difficult. For example, there are a lot of abbreviations that were not previously defined and/or are not specified if they are genes, proteins operons or something similar. Finally, I think a perspective section, before the conclusion, is necessary for this manuscript. check the reference list according to the MDPI format. Here are my specific comments and suggestions that authors must work in order to improve the scientific quality of the manuscript.
If possible, give more details about this “crucial role”
Abstract:
Line 10: Is there a particular species causing brucellosis? Or do all the genera in brucella cause brucellosis? Add this info for readers that does know a lot about brucellosis
Line 12: provide a short definition about the function of the virB system. Is it an operon? Involved in what?
Keywords:
Line 20: The keywords can be improved
Section 1:
Line 28: among others? A sentence explaining that brucella spp. cause brucellosis is necessary. Define if there is an specific genera involved in this disease
Line 32: A process THATdepends?
Line 33: a paragraph summarizing the main mechanisms revealed is necessary for better contextualization in this section
Section 2:
Line 38: operon
Line 39: delete “(B. suis)”
Line 41:what is “orfs”
Line 43: reference number missing for Hartigh
Line 49: please rewrite to “(Table 1)”
Line 50: references missing in table 1
Line 53: other
Line 54 please rewrite to: (Fig. 1) the same for all figures and tables
Line 56: Please. Provide a figure caption, not literature review or discussion
Line 64: improve the title of this section. The same al line 71, 89, and 96
Line 68: reference missing
Line 73: reference missing
Line 74: reference missing
Line 79: reference missing
Paragraph in in line 64 to 105: please refer to the fig.1 wherever necessary
Line 86: I do not think this abbreviation is necessary in this manuscript (“OM”) as the authors have lots of abbreviations. the same for IM, TM and abbreviations like that. Please check
Line 91: reference missing
Line 97: change , by .
Section 3:
Line 109: What is IMC and OMC? It was defined previously?
Line 115 to 118: this sentence must be rewritten
Line 121: a new sentence must be started after “aggregation”
Line 126: delete “,”
Paragraph at line 108: a diagram or figure explaining those processes will benefits the understanding of the detailed information
Line 136: “Second”. Change “is” by “must be”
Sentence at line 135: are references for this? More information must be added to this section. I feel that is too short
Line 138: was RicA previously defined?
Line 140: reference missing
Line 142: reference missing
Line 142: delete “in” and “, it”
Line 143: was “LBH” previously defined?
Line 145:what is rab2
Line 146: what is igfp
Line 147: What is gdp?
Line 148: what is vcec and veca?
Line 156: what is jbr, p62, lc3, bcl, and many other abbreviations in the manuscript that must be defined
Line 157: Reference number missing for zhang et al
Line 163: I feel that in this paragraph the authors have only described the available literature, without a clear discussion made by the authors. The same thing for the others paragraphs present in the section 3
Line 167: Add an space after 250
Section 4:
Line : 188: it should be Brucella?
Line 193: correct the “,” and Brucella
Line 193: what “eres” is?
Figure 2: clearly indicate where brucella is
Line 212: reference number missing in celli et al
Line 215: showing?
Line 222 to 225: i feel that this sentence must be rewritten or separated in two
Line 227: reference number missing for smith et al
Figure 3: Please, maintain the format of the previous figure (white background) the resolution of all figures must be enhanced. Similarly, figure caption can be improved considering only description of the thing present in the figure
Line: Unfolded in lowercase
Line 251: reference number missing for yuki et al
Line 256: please. Merge or separate both sentences “. And”
Line 258 What is CHOP ?
Line 268: Mal in uppercase
Line 269: delete obviously
Line 273: in-vitro in italics
Line 293: deletes spaces before “,”
Line 299: reference missing
Line 302: Kinases
Line 320: delete “.” before “and”. Additionally, the sentence must be rewritten. It is confusing
Line 336: a protein THAT negatively ?
Sentence at line 351: this sentenced need to be shortened or separated in two or three
Author Response
Cover Letter for revised version of ijms-1470404
Dear Drs. Madam and/or Sir
I am writing the Cover Letter to submit our revised manuscript entitled, “The VirB system plays a crucial role in Brucella intracellular infection” for publication in International Journal of Molecular Sciences. Our research group made our best effort to address all comments/suggestions from three reviewers. We believe the revised version could be ready to be considered for publication in your journal. All authors approved the revised version and this submission.
Thank you for receiving our revised manuscript and considering it for publication. We appreciate your time and look forward to hearing your response.
Sincerely,
Hanwei Jiao, DVM, PhD
Abstract:
Line 10: Is there a particular species causing brucellosis? Or do all the genera in brucella cause brucellosis? Add this info for readers that does know a lot about brucellosis
Response: Thank you for your comments. All the genera in brucella can cause brucellosis. In line10, we have changed “Brucellosis is a highly prevalent zoonotic disease caused by Brucella” to “Brucellosis is a highly prevalent zoonotic disease caused by Brucella spp.”.
Line 12: provide a short definition about the function of the virB system. Is it an operon? Involved in what?
Response: We really appreciate your constructive proposal, I have changed“depend on a functional virB system”to “ depend on a functional virB system, an operon encoded by VirB1-VirB12”
Keywords:
Line 20: The keywords can be improved
Response: Thank you very much for your suggestion. We have changed the keywords “VirB, T4SS” to “VirB system, type IV secretion”
Section 1:
Line 28: among others? A sentence explaining that brucella spp. cause brucellosis is necessary. Define if there is an specific genera involved in this disease
Response: I really appreciate your constructive proposal, here’s what we added to line28 “Brucella species frequently isolated from job-related infection were Brucella melitensis, Brucella abortus, Brucella suis and Brucella canis.”
Line 32: A process THATdepends?
Response: I’m sorry I didn’t make myself clear, we have changed “a process depends on T4SS encoded by VirB operon” to “a intracellular process depends on type IV secretion which encoded by VirB operon. ”
Line 33: a paragraph summarizing the main mechanisms revealed is necessary for better contextualization in this section
Response: I really appreciate your constructive proposal, we have added the following “During intracellular infection, VirB T4SS is rapidly activated and reaches its maximum activity in five hours, when the replication niche is established, VirB T4SS is inhibited. During this period，VirB T4SS can subvert the cell pathway of host cells, which is beneficial to the bacteria reproduction and pathogenicity.”
Section 2:
Line 38: operon
Response: Thank you for your careful reading. We have corrected this word.
Line 39: delete “(B. suis)”
Response: Thank you very much for your suggestion. We deleted “(B. suis)”.
Line 41:what is “orfs”
Response: I’m sorry I didn’t make myself clear, “orfs” is open reading frames, I have changed “orfs” to “open reading frames (ORFs)”.
Line 43: reference number missing for Hartigh
Response: Thank you for your correction. We have added the reference.
Line 49: please rewrite to “(Table 1)”
Response: Thank you for your correction. We have changed “Table 1” to “(Table 1)”.
Line 50: references missing in table 1
Response: Thank you for your correction. We have added references in table 1.
Line 53: other
Response: Thank you for your correction. We have changed “Other” to “other”.
Line 54 please rewrite to: (Fig. 1) the same for all figures and tables
Response: Thank you for your correction. We have rewritten “Fig. 1” to “(Fig. 1)”, the same for all figures and tables.
Line 56: Please. Provide a figure caption, not literature review or discussion
Response: Thank you very much for your suggestion. We have changed the captions to “ Fig. 1. VirB system structure. The prototype of Brucella VirB system is Agrobacterium tumefaciens VirB/D4 system. The VirB system of Brucella is composed of VirB1-VirB12 and divided into five parts according to the structure: the inner membrane area is composed of VirB3, VirB4, VirB6, VirB8 and VirB10; the central and adventitial areas include VirB7, VirB9 and VirB10; the junction area is composed of VirB5 and VirB10; the extension consists of VirB2; the energy supply area is composed of VirB4 and VirB11. Present, it is not clear the exact location of VirB11 relative to VirB4, it can accumulate under the VirB4 or on one side of the cytoplasm.”
Line 64: improve the title of this section. The same al line 71, 89, and 96
Response: Thank you very much for your suggestion. We have changed “Core component” to “Core component VirB6-VirB10”; “The surface exposure components” to “ The surface exposure components VirB2 and VirB5”; “Other components” to “Other components VirB1 and VirB12”
Line 68: reference missing
Response: Thank you for your correction. We have added the reference.
Line 73: reference missing
Response: Thank you for your correction. We have added the reference.
Line 74: reference missing
Response: Thank you for your correction. We have added the reference.
Line 79: reference missing
Response: Thank you for your correction. We have added the reference.
Paragraph in in line 64 to 105: please refer to the fig.1 wherever necessary
Response: We really appreciate your constructive proposal, We have made the changes as you suggested.
Line 86: I do not think this abbreviation is necessary in this manuscript (“OM”) as the authors have lots of abbreviations. the same for IM, TM and abbreviations like that. Please check
Response: We really appreciate your constructive proposal, We have changed “TM” to “transmembrane”, “IM” to “inner membrane”, “NTD” to “N-terminal domain”; “CTD” changed to “C-terminal domain”, “OM” to “outer membrane”.
Line 91: reference missing
Response: Thank you for your correction. We have added the reference.
Line 97: change , by .
Response: Thank you very much for your suggestion. We have changed “,” by “.”
Section 3:
Line 109: What is IMC and OMC? It was defined previously?
Response: Thank you for your correction. It was not defined previously. OMC is a large structure inserted into the inner membrane and similar to the core complex. OMC is connected by a flexible central stalk to a larger complex, the intimal complex (IMC)
Line 115 to 118: this sentence must be rewritten
Response: Thank you very much for your suggestion. We have rewritten “In the three basic parts of T4SS, the core complex connects IM and OM, acts as the outer membrane component of the substrate transmembrane transport channel in the periplasm，the inner membrane component of this channel is composed of VirB3, VirB6 and VirB8” as“In the basic structure of T4SS, the substrate transmembrane transport channel in the periplasm connects the outer membrane and the inner membrane. The inner membrane of the channel is composed of VirB3, VirB6 and VirB8”.
Line 121: a new sentence must be started after “aggregation”
Response: Thank you very much for your suggestion. We have added “and then VirB9 dissociated VirB8−VirB10 dimer”
Line 126: delete “,”
Response: Thank you very much for your suggestion. We have deleted “,”.
Paragraph at line 108: a diagram or figure explaining those processes will benefits the understanding of the detailed information
Response: Thank you very much for your suggestion. We checked the relevant References:
[1]Ward DV, Draper O, Zupan JR, Zambryski PC. Peptide linkage mapping of the Agrobacterium tumefaciens vir-encoded type IV secretion system reveals protein subassemblies. Proceedings of the National Academy of Sciences of the United States of America 2002ï¼›99:11493-11500
[2] Carle A, Hoppner C, Ahmed Aly K, Yuan Q, den Dulk-Ras A, Vergunst A, O'Callaghan D, Baron C. The Brucella suis type IV secretion system assembles in the cell envelope of the heterologous host Agrobacterium tumefaciens and increases IncQ plasmid pLS1 recipient competence. Infect Immun 2006ï¼›74:108-17.
[3] Cascales E, Christie PJ. Agrobacterium VirB10, an ATP energy sensor required for type IV secretion. Proc Natl Acad Sci U S A 2004ï¼›101:17228-33.
[4] Yuan Q, Carle A, Gao C, Sivanesan D, Aly KA, Hoppner C, Krall L, Domke N, Baron C. Identification of the VirB4-VirB8-VirB5-VirB2 pilus assembly sequence of type IV secretion systems. J Biol Chem 2005ï¼›280:26349-59.
[5] Hoppner C, Carle A, Sivanesan D, Hoeppner S,Baron C. The putative lytic transglycosylase VirB1 from Brucella suis interacts with the type IV secretion system core components VirB8, VirB9 and VirB11. Microbiology (Reading) 2005ï¼›151:3469-3482.
There is no systematic description of the formation process of T4SS. The formation of T4SS remains to be studied. In addition, the formation of T4SS is a dynamic process, which can’t be described by a few pictures.
Line 136: “Second”. Change “is” by “must be”
Response: Thank you very much for your suggestion. We have modified “second” by “Second”, changed “is” by “must be”.
Sentence at line 135: are references for this? More information must be added to this section. I feel that is too short
Response: Yes, the references are for this. We have added some information. Here’s what’s changed “Although T4SS was discovered in Brucella spp. about 15 years ago, the T4SS effectors were confirmed until recently. A candidate protein can be identified as a T4SS effector protein if it meets two criteria: first, the protein must be secreted by VirB T4SS. Second, this protein must be able to enter the host cell [7,8]. The first criterion can be verified by constructing T4SS defective strain, the second standard can be verified by TEM-1 lactamase or calmodulin-dependent adenylate cyclase assays [8]. There are currently 15 known VirB T4SS effectors: RicA, VceC, VecA, BtpA, BtpB, BspA, BspB, BspF, BPE005, BPE123, BPE043, BPE275, SepA, BspC, and BspE, they manipulate host cell pathways and responses [7,8].”
Line 138: was RicA previously defined?
Response: I am sorry. It wasn’t previously defined. RicA is an effector of T4SS. I’ve added that to the paper. “There are currently 15 known VirB T4SS effectors: RicA, VceC, VecA, BtpA, BtpB, BspA, BspB, BspF, BPE005, BPE123, BPE043, BPE275, SepA, BspC, and BspE.”
Line 140: reference missing
Response: Thank you for your correction. We have added the reference.
Line 142: reference missing
Response: Thank you for your correction. We have added the reference.
Line 142: delete “in” and “, it”
Response: Thank you very much for your suggestion. We have deleted “in” and “, it”.
Line 143: was “LBH” previously defined?
Response: I am sorry. It wasn’t previously defined. LβH is a protein superfamily including acetyltransferases, ferripyochelin binding proteins, acyltransferases, carbonic anhydrases, and some proteins of unknown functions.
Line 145:what is rab2
Response: Rab2 is a cellular host factor critical for brucella intracellular proliferation.
Line 146: what is igfp
Response: The isoleucine-glycine-phenylalanine-proline (IGFP) loop is part of a sequence of 8 amino acids in length flanked on each side by a β strand of the LβH.
Line 147: What is gdp?
Response: GDP is Guanosine diphosphate.
Line 148: what is vcec and veca?
Response: They are T4SS effectors.
Line 156: what is jbr, p62, lc3, bcl, and many other abbreviations in the manuscript that must be defined
Response: VjbR is a quorum sensing regulatory protein. BCL-2 is an anti-apoptotic gene. p62, LC3-II, Atg5 and Caspase-3 are autophagy genes. We have made some changes to the above.
Line 157: Reference number missing for zhang et al
Response: Thank you for your correction. We have added the reference.
Line 163: I feel that in this paragraph the authors have only described the available literature, without a clear discussion made by the authors. The same thing for the others paragraphs present in the section 3
Response: Thank you very much for your suggestion. We have added a discussion in section 3.
Line 167: Add an space after 250
Response: Thank you for your correction. We have added an space after 250.
Section 4:
Line : 188: it should be Brucella?
Response: Thank you for your correction. We have changed it by “Brucella”.
Line 193: correct the “,” and Brucella
Response: Thank you for your correction. We have changed the “,” and Brucella.
Line 193: what “eres” is?
Response: ERES is endoplasmic reticulum exit sites (ERES).
Figure 2: clearly indicate where brucella is
Response: Thank you very much for your suggestion. We have made changes to the Figure 2
Line 212: reference number missing in celli et al
Response: Thank you for your correction. We have added the reference.
Line 215: showing?
Response: Thank you for your correction. We have changed “show” by “showing”.
Line 222 to 225: i feel that this sentence must be rewritten or separated in two
Response: Thank you very much for your suggestion. This sentence has been separated in two.
Line 227: reference number missing for smith et al
Response: Thank you for your correction. We have added the reference.
Figure 3: Please, maintain the format of the previous figure (white background) the resolution of all figures must be enhanced. Similarly, figure caption can be improved considering only description of the thing present in the figure
Response: Thank you very much for your suggestion. We have made the changes as you suggested
Line: Unfolded in lowercase
Response: Thank you for your correction. We have changed it by “unfolded”.
Line 251: reference number missing for yuki et al
Response: Thank you for your correction. We have added the reference.
Line 256: please. Merge or separate both sentences “. And”
Response: Thank you very much for your suggestion. We have separated it.
Line 258 What is CHOP ?
Response: CHOP is C/EBP-homologous protein.
Line 268: Mal in uppercase
Response: Thank you very much for your suggestion. We have changed it.
Line 269: delete obviously
Response: Thank you very much for your suggestion. We have deleted “obviously”.
Line 273: in-vitro in italics
Response: Thank you for your correction. We have changed it in italics.
Line 293: deletes spaces before “,”
Response: Thank you for your correction. We have deleted it.
Line 299: reference missing
Response: Thank you for your correction. We have added references.
Line 302: Kinases
Response: Thank you for your correction. We have changed it by “Kinases”
Line 320: delete “.” before “and”. Additionally, the sentence must be rewritten. It is confusing
Response: Thank you very much for your suggestion. Here’s what We changed “ The transcriptional repressor of the histidine utilization (hut) genes HutC directly controls the activity of the virB promoter (PvirB) Under the specific condition of PH4.5. This regulation is related to the induction of histidine utilization pathway and the regulation may be a way to evade host immunity by perceiving self-metabolism and developing an adaptive response [65].”
Line 336: a protein THAT negatively ?
Response: Thank you for asking. We want to express is not a negative protein, but this protein inhibits other functions.
Sentence at line 351: this sentenced need to be shortened or separated in two or three
Response: Thank you very much for your suggestion. This sentence has been divided separated in two
Reviewer 2 Report
The manuscript “The VirB system plays a crucial role in Brucella intracellular infection” Xue Xiong et all. is a review on brucella VirB system and its function during infection. The authors described what is known about the mechanisms and proteins encoded by VirB operon. In addition the Authors summarize the intacelullar brucella circulation and the consequences for some cellular pathways related to brucella infection. In this review a recent knowledge is present and in my opinion this manuscript updates previously published reviews, therefore it might be of interest for readers working in the field of microbiology. However, the manuscript requires extensive spellcheck and rephrasing of some parts to avoid misunderstanding and to provide high quality and biological correct information.
As for example:
L40 13 orfs or 12 orfs?
L42 a operon – an operon
L53 and Other – and other
L56 Fig. 1. VirB operon structure. – misleading as this is not a structure of the operon
L65-70 needs rephrasing
L80-83 needs rephrasing to make more clear what is consisting of a n-terminal domain……
L189 Brucella invades host cells through inhalation……. – should be rephrased, as a cell cannot uptake a bacteria through inhalation and so on.
And many more.
Author Response
Cover Letter for revised version of ijms-1470404
Dear Drs. Madam and/or Sir
I am writing the Cover Letter to submit our revised manuscript entitled, “The VirB system plays a crucial role in Brucella intracellular infection” for publication in International Journal of Molecular Sciences. Our research group made our best effort to address all comments/suggestions from three reviewers. We believe the revised version could be ready to be considered for publication in your journal. All authors approved the revised version and this submission.
Thank you for receiving our revised manuscript and considering it for publication. We appreciate your time and look forward to hearing your response.
Sincerely,
Hanwei Jiao, DVM, PhD
Question 1：L40 13 orfs or 12 orfs?
Response: As you suggested, we checked "L40 13 orfs" and modified the error here to "12 ofrs". We thank you very much for your suggestion.
Question 2：L42 a operon – an operon
Response: As you suggested, we made corresponding changes in the revised manuscript R1 and marked it in blue.
Question 3：L53 and Other – and other
Response: As you suggested, we made corresponding changes in the manuscript and marked it in blue.
Question 4：L56 Fig. 1. VirB operon structure. – misleading as this is not a structure of the operon
Response: Thank you for your correction. We have corrected it to “VirB system structure”.
Question 5：L65-70 needs rephrasing
Response: As you suggested, we reworded lines 68-72 (L65-70 in original version) of the revised manuscript R1 and marked them in blue.
Question 6：L80-83 needs rephrasing to make more clear what is consisting of a n-terminal domain……
Response: Response: This sentence of L82-88 in the revised manuscript R1 (L80-83 in original version) has been rephrased according to your suggestions as follows: VirB10 can pass through the entire outer membrane of Brucella T4SS structure. Therefore, VirB10 is an important functional protein for Brucella. The protein consists of an N-terminal domain, it is found that the domain contains four parts: a cytoplasmic component, a TM helix, a bendable component and a spherical CTD. These four parts lead to the central function of VirB10 protein in T4SS, that is, connecting many different proteins and sending interactions, so as to mediate the signal transmission process.
Question 7：L189 Brucella invades host cells through inhalation……. – should be rephrased, as a cell cannot uptake a bacteria through inhalation and so on.
Response: Response: As you suggested, we reorganized this sentence of L212 in the revised manuscript R1 (L189 in original version: Brucella can invade the body through skin mucosa, digestive tract and respiratory tract, then it is enclosed in a membrane to form a Brucella-containing vacuole (BCV) [54]) according to your request.
Question 8: And many more.
Response: Some writing problems were pointed out by other reviewers, revised and marked in the revised manuscipt R1. As you suggested, We re-examined this manuscript very carefully, and reworded some descriptions and marked them in blue. The specific modifications are as follows:
- L49-50 in revised manuscript R1 (Facts have proved that VirB2 plays an important role in the continuous infection of mice caused by Brucella, but VirBl is dispensable [12].)
- L108-110 in revised manuscript R1 (The exact location of VirBl2 is still unclear, it is only confirmed that it is a cell surface protein that induces antibody response during animal infection.)
- L337-340 in revised manuscript R1 (The expression of VirB system peaked during rBCV biogenesis, but the bacteria gradually began to.....
- L341-346 in revised manuscript R1 (Factors that directly regulate the VirB system includeHutc, IFH, quorum sensing regulatory protein BlxR and autoinducers in the quorum)
We have tried our best to find the inappropriate places in the article, and invited a native English speaking experts to modify the language. If there are still mistakes, we strongly hope to get your advices, and we will modify them.
Reviewer 3 Report
This review summarizes the mechanism of VirB action by showing the function and the significance of VirB system, focusing on the structure of virB system, VirB T4SS mediates biogenesis of the endoplasmic reticulum (ER)-derived replicative Brucella-containing vacuole (rBCV), the effectors of T4SS and the cellular pathways it subverted. This aims to provide a theoretical basis for the pathogenesis of Brucella in order to provide new ideas for the research and development of clinical vaccines. This paper is interesting and deserves to be published after a minor revision.
The corrections are detailed below:
Line 10: Please change “Brucella is a gram” by “Brucella spp. are gram”.
Line 14: Change “which is conducive to the replication of Brucella” by “which promotes Brucella replication”.
Line 18: Delete “to” before “better”.
Line 19: Change “for the research and development of clinical vaccines.” by “for clinical vaccine research and development.”.
Line 25: Modify “which brings great economic losses to the animal husbandry industry [2].” by “resulting in great economic losses to the livestock husbandry industry [2].”
Line 27: Change “or by eating contaminated dairy products, with clinical symptoms such as intermittent fever, arthritis, orchitis, hepatitis, depression [2,3].” by “or by consumption of contaminated dairy products, with clinical symptoms such as intermittent fever, arthritis, orchitis, hepatitis, and depression [2,3].”.
Line 32: Modify “process depends” by “process dependent”.
Line 34: Change “circulation largely has been elucidated” by “circulation has been largely elucidated”.
Line 40: Change “all Brucella pecies” by “all Brucella species”
Line 94: Add “to” before “integrate”.
Line 102: Delete “of” before “bacterial cells”.
Line 121: Change “separate” by “separates”.
Line 136: Modify “second” by “Second”.
Line 140: Change “consisting” by “consists”.
Line 144: Modify “is exposed to the exterior [38-40].” by “is externally exposed [38-40].”.
Lines 165 and 166: Change “brucella” by “Brucella”.
Line 188: Change “bercella” by “Brucella”.
Line 193: Change “Immediately after , brucella interacted” by “Immediately after, Brucella interacts”.
Lines 212 and 260: Modify “infection of Brucella” by “Brucella infection”.
Line 222: Change “to establish replication niche” by “to establish a replication niche”.
Line 236: Add “the” before “virulence”.
Line 256: Change “the inflammation [68]. And when the ER…” by “the inflammation [68], and when the ER…”.
Line 266: Change “to decreases” by “to decrease”.
Line 271: Modify “funcions” by “functions”.
Line 272: Change “by the way of inhibiting the transduction” by “by inhibiting transduction”.
Line 284: Modify “In the aspect of biogenesis of rBVC” by “In the aspect of rBVC biogenesis”.
Line 285: Change “of host early secretory pathway” by “of host’s early secretory pathway”.
Line 297: Modify “T4SS may secretes effector…” by “T4SS may secretes an effector…”.
Line 302 Change “kinases” by “Kinases”.
Line 304: Modify “It should also be noted that” by “It is also worth noting that”.
Line 307: Change “between the ER and Golgi” by “between the ER and the Golgi apparatus”.
Line 321: Modify “induced on the later stage of infection” by “induced at the later stage of infection”.
Author Response
Cover Letter for revised version of ijms-1470404
Dear Drs. Madam and/or Sir
I am writing the Cover Letter to submit our revised manuscript entitled, “The VirB system plays a crucial role in Brucella intracellular infection” for publication in International Journal of Molecular Sciences. Our research group made our best effort to address all comments/suggestions from three reviewers. We believe the revised version could be ready to be considered for publication in your journal. All authors approved the revised version and this submission.
Thank you for receiving our revised manuscript and considering it for publication. We appreciate your time and look forward to hearing your response.
Sincerely,
Hanwei Jiao, DVM, PhD
The corrections are detailed below:
Line 10: Please change “Brucella is a gram” by “Brucella spp. are gram”.
Response: Thank you very much for your suggestion. We have changed “Brucella is a gram” to “Brucella spp. are gram”.
Line 14: Change “which is conducive to the replication of Brucella” by “which promotes Brucella replication”.
Response: We have changed “which is conducive to the replication of Brucella” by “which promotes Brucella replication”.
Line 18: Delete “to” before “better”.
Response: We have deleted “to” before “better”.
Line 19: Change “for the research and development of clinical vaccines.” by “for clinical vaccine research and development.”.
Response: We have changed “for the research and development of clinical vaccines.” by “for clinical vaccine research and development.”.
Line 25: Modify “which brings great economic losses to the animal husbandry industry [2].” by “resulting in great economic losses to the livestock husbandry industry [2].”
Response: We have modified “which brings great economic losses to the animal husbandry industry [2].” by “resulting in great economic losses to the livestock husbandry industry [2].”
Line 27: Change “or by eating contaminated dairy products, with clinical symptoms such as intermittent fever, arthritis, orchitis, hepatitis, depression [2,3].” by “or by consumption of contaminated dairy products, with clinical symptoms such as intermittent fever, arthritis, orchitis, hepatitis, and depression [2,3].”.
Response: We have changed “or by eating contaminated dairy products, with clinical symptoms such as intermittent fever, arthritis, orchitis, hepatitis, depression [2,3].” by “or by consumption of contaminated dairy products, with clinical symptoms such as intermittent fever, arthritis, orchitis, hepatitis, and depression [2,3].”.
Line 32: Modify “process depends” by “process dependent”.
Response: We have modified “process depends” by “process dependent”.
Line 34: Change “circulation largely has been elucidated” by “circulation has been largely elucidated”.
Response: We have changed “circulation largely has been elucidated” by “circulation has been largely elucidated”.
Line 40: Change “all Brucella pecies” by “all Brucella species”
Response: We have changed “all Brucella pecies” by “all Brucella species”.
Line 94: Add “to” before “integrate”.
Response: We have added “to” before “integrate”.
Line 102: Delete “of” before “bacterial cells”.
Response: We have deleted “of” before “bacterial cells”.
Line 121: Change “separate” by “separates”.
Response: We have changed “separate” by “separates”.
Line 136: Modify “second” by “Second”.
Response: We have modified “second” by “Second”.
Line 140: Change “consisting” by “consists”.
Response: We have changed “consisting” by “consists”.
Line 144: Modify “is exposed to the exterior [38-40].” by “is externally exposed [38-40].”.
Response: We have modified “is exposed to the exterior [38-40].” by “is externally exposed [38-40].”.
Lines 165 and 166: Change “brucella” by “Brucella”.
Response: We have changed “brucella” by “Brucella”.
Line 188: Change “bercella” by “Brucella”.
Response: We have changed “bercella” by “Brucella”.
Line 193: Change “Immediately after , brucella interacted” by “Immediately after, Brucella interacts”.
Response: We have changed “Immediately after , brucella interacted” by “Immediately after, Brucella interacts”.
Lines 212 and 260: Modify “infection of Brucella” by “Brucella infection”.
Response: We have modified “infection of Brucella” by “Brucella infection”.
Line 222: Change “to establish replication niche” by “to establish a replication niche”.
Response: We have changed “to establish replication niche” by “to establish a replication niche”.
Line 236: Add “the” before “virulence”.
Response: We have added “the” before “virulence”.
Line 256: Change “the inflammation [68]. And when the ER…” by “the inflammation [68], and when the ER…”.
Response: We have changed “the inflammation [68]. And when the ER…” by “the inflammation [68], and when the ER…”.
Line 266: Change “to decreases” by “to decrease”.
Response: We have changed “to decreases” by “to decrease”.
Line 271: Modify “funcions” by “functions”.
Response: We have modified “funcions” by “functions”.
Line 272: Change “by the way of inhibiting the transduction” by “by inhibiting transduction”.
Response: We have changed “by the way of inhibiting the transduction” by “by inhibiting transduction”.
Line 284: Modify “In the aspect of biogenesis of rBVC” by “In the aspect of rBVC biogenesis”.
Response: We have modified “In the aspect of biogenesis of rBVC” by “In the aspect of rBVC biogenesis”.
Line 285: Change “of host early secretory pathway” by “of host’s early secretory pathway”.
Response: We have changed of host early secretory pathway” by “of host’s early secretory pathway”.
Line 297: Modify “T4SS may secretes effector…” by “T4SS may secretes an effector…”.
Response: We have modified “T4SS may secretes effector…” by “T4SS may secretes an effector…”.
Line 302 Change “kinases” by “Kinases”.
Response: We have changed “kinases” by “Kinases”.
Line 304: Modify “It should also be noted that” by “It is also worth noting that”.
Response: We have modified “It should also be noted that” by “It is also worth noting that”.
Line 307: Change “between the ER and Golgi” by “between the ER and the Golgi apparatus”.
Response: We have changed “between the ER and Golgi” by “between the ER and the Golgi apparatus”.
Line 321: Modify “induced on the later stage of infection” by “induced at the later stage of infection”.
Response: We have modified “induced on the later stage of infection” by “induced at the later stage of infection”.
Round 2
Reviewer 1 Report
the manuscript have been improved in accordance with my previous comments. I have no further suggestion
Author Response
Thank you for your constructive and detailed comments, which substantially improved the quality of our manuscript.
Reviewer 2 Report
The Authors made a good job, but still some minor revision should be made:
L89-91: “The protein consists of an N-terminal domain, it is found that the domain contains four parts: a cytoplasmic component, a TM helix, a bendable component and a spherical CTD” – in my opinion it is not right, the spherical CTD is not a part of NTD. Or CTD stays not for C-terminal domain ?
Author Response
Thank you for your question. We made our best effort to address your comments. L89-91 were actually quoted in this reference:
[16] Berge C, Waksman G,Terradot L. Structural and Molecular Biology of Type IV Secretion Systems. Curr Top Microbiol Immunol 2017ï¼›413:31-60.
L89-91 stated in the original reference as:
“VirB10 is a remarkable protein since it traverses the entire bacterial envelope in Gram-bacterial T4SSs. The protein is composed of an N-terminal domain that contains a small cytoplasmic portion, a TM helix, a flexible part, and a globular CTD.” It shows that the spherical CTD is a part of NTD.
