KCTD Proteins Have Redundant Functions in Controlling Cellular Growth
Abstract
:1. Introduction
2. Results
2.1. Transcriptional Profiles Associated with the Decrease in Cell Growth KCTD-KO Cells
2.2. Impact of KCTD and GNB1 Knockouts on G Protein Transcriptional Profiles
2.3. Calcium Signalling and KCTD Isoforms
3. Discussion
4. Materials and Methods
4.1. Sequence Alignment of KCTD Proteins
4.2. Generation of Knockout Lines
4.3. Immunodetection of Gβ1
4.4. Cell Growth Assays
4.5. Calcium Measurements
4.6. RNA Extraction, Library Preparation and Sequencing
4.7. Bioinformatic Analysis
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Smaldone, G.; Coppola, L.; Incoronato, M.; Parasole, R.; Ripaldi, M.; Vitagliano, L.; Mirabelli, P.; Salvatore, M. KCTD15 Protein Expression in Peripheral Blood and Acute Myeloid Leukemia. Diagnostics 2020, 10, 371. [Google Scholar] [CrossRef] [PubMed]
- Angrisani, A.; Di Fiore, A.; De Smaele, E.; Moretti, M. The emerging role of the KCTD proteins in cancer. Cell Commun. Signal. 2021, 19, 56. [Google Scholar] [CrossRef]
- Liu, Z.; Xiang, Y.; Sun, G. The KCTD family of proteins: Structure, function, disease relevance. Cell Biosci. 2013, 3, 45. [Google Scholar] [CrossRef] [PubMed]
- Yao, H.; Ren, D.; Wang, Y.; Wu, L.; Wu, Y.; Wang, W.; Li, Q.; Liu, L. KCTD9 inhibits the Wnt/β-catenin pathway by decreasing the level of β-catenin in colorectal cancer. Cell Death Dis. 2022, 13, 761. [Google Scholar] [CrossRef]
- Teng, X.; Aouacheria, A.; Lionnard, L.; Metz, K.A.; Soane, L.; Kamiya, A.; Hardwick, J.M. KCTD: A new gene family involved in neurodevelopmental and neuropsychiatric disorders. CNS Neurosci. Ther. 2019, 25, 887–902. [Google Scholar] [CrossRef]
- Ahmed, S.M.; Daulat, A.M.; Meunier, A.; Angers, S. G protein βγ subunits regulate cell adhesion through Rap1a and its effector Radil. J. Biol. Chem. 2010, 285, 6538–6551. [Google Scholar] [CrossRef]
- Campden, R.; Petrin, D.; Robitaille, M.; Audet, N.; Gora, S.; Angers, S.; Hebert, T.E. Tandem affinity purification to identify cytosolic and nuclear gβγ-interacting proteins. Methods Mol. Biol. 2015, 1234, 161–184. [Google Scholar] [CrossRef] [PubMed]
- Young, B.D.; Sha, J.; Vashisht, A.A.; Wohlschlegel, J.A. Human Multisubunit E3 Ubiquitin Ligase Required for Heterotrimeric G-Protein β-Subunit Ubiquitination and Downstream Signaling. J. Proteome Res. 2021, 20, 4318–4330. [Google Scholar] [CrossRef]
- Muntean, B.S.; Marwari, S.; Li, X.; Sloan, D.C.; Young, B.D.; Wohlschlegel, J.A.; Martemyanov, K.A. Members of the KCTD family are major regulators of cAMP signaling. Proc. Natl. Acad. Sci. USA 2022, 119, e2119237119. [Google Scholar] [CrossRef]
- Brockmann, M.; Blomen, V.A.; Nieuwenhuis, J.; Stickel, E.; Raaben, M.; Bleijerveld, O.B.; Altelaar, A.F.M.; Jae, L.T.; Brummelkamp, T.R. Genetic wiring maps of single-cell protein states reveal an off-switch for GPCR signalling. Nature 2017, 546, 307–311. [Google Scholar] [CrossRef]
- Sloan, D.C.; Cryan, C.E.; Muntean, B.S. Multiple potassium channel tetramerization domain (KCTD) family members interact with Gβγ, with effects on cAMP signaling. J. Biol. Chem. 2023, 299, 102924. [Google Scholar] [CrossRef] [PubMed]
- Fritzius, T.; Turecek, R.; Seddik, R.; Kobayashi, H.; Tiao, J.; Rem, P.D.; Metz, M.; Kralikova, M.; Bouvier, M.; Gassmann, M.; et al. KCTD Hetero-oligomers Confer Unique Kinetic Properties on Hippocampal GABAB Receptor-Induced K+ Currents. J. Neurosci. 2017, 37, 1162–1175. [Google Scholar] [CrossRef]
- Skoblov, M.; Marakhonov, A.; Marakasova, E.; Guskova, A.; Chandhoke, V.; Birerdinc, A.; Baranova, A. Protein partners of KCTD proteins provide insights about their functional roles in cell differentiation and vertebrate development. Bioessays 2013, 35, 586–596. [Google Scholar] [CrossRef]
- Nguyen, D.M.; Rath, D.H.; Devost, D.; Petrin, D.; Rizk, R.; Ji, A.X.; Narayanan, N.; Yong, D.; Zhai, A.; Kuntz, D.A.; et al. Structure and dynamics of a pentameric KCTD5/CUL3/Gβγ E3 ubiquitin ligase complex. Proc. Natl. Acad. Sci. USA 2024, 121, e2315018121. [Google Scholar] [CrossRef] [PubMed]
- Ji, A.X.; Chu, A.; Nielsen, T.K.; Benlekbir, S.; Rubinstein, J.L.; Prive, G.G. Structural Insights into KCTD Protein Assembly and Cullin3 Recognition. J. Mol. Biol. 2016, 428, 92–107. [Google Scholar] [CrossRef]
- Pirone, L.; Correale, S.; de Paola, I.; Zaccaro, L.; De Simone, G.; Vitagliano, L.; Pedone, E.; Di Gaetano, S. Design, synthesis and characterization of a peptide able to bind proteins of the KCTD family: Implications for KCTD-cullin 3 recognition. J. Pept. Sci. 2011, 17, 373–376. [Google Scholar] [CrossRef] [PubMed]
- Jiang, W.; Wang, W.; Kong, Y.; Zheng, S. Structural basis for the ubiquitination of G protein βγ subunits by KCTD5/Cullin3 E3 ligase. Sci. Adv. 2023, 9, eadg8369. [Google Scholar] [CrossRef]
- Khan, S.M.; Martin, R.D.; Bayne, A.; Petrin, D.; Bourque, K.; Jones-Tabah, J.; Bouazza, C.; Blaney, J.; Lau, J.; Martins-Cannavino, K.; et al. Gβγ subunits colocalize with RNA polymerase II and regulate transcription in cardiac fibroblasts. J. Biol. Chem. 2023, 299, 103064. [Google Scholar] [CrossRef]
- Khan, S.M.; Min, A.; Gora, S.; Houranieh, G.M.; Campden, R.; Robitaille, M.; Trieu, P.; Petrin, D.; Jacobi, A.M.; Behlke, M.A.; et al. Gβ4γ1 as a modulator of M3 muscarinic receptor signalling and novel roles of Gβ1 subunits in the modulation of cellular signalling. Cell. Signal. 2015, 27, 1597–1608. [Google Scholar] [CrossRef]
- Robitaille, M.; Gora, S.; Wang, Y.; Goupil, E.; Petrin, D.; Del Duca, D.; Villeneuve, L.R.; Allen, B.G.; Laporte, S.A.; Bernard, D.J.; et al. Gβγ is a negative regulator of AP-1 mediated transcription. Cell. Signal. 2010, 22, 1254–1266. [Google Scholar] [CrossRef]
- Khan, S.M.; Sleno, R.; Gora, S.; Zylbergold, P.; Laverdure, J.P.; Labbe, J.C.; Miller, G.J.; Hebert, T.E. The expanding roles of Gβγ subunits in G protein-coupled receptor signaling and drug action. Pharmacol. Rev. 2013, 65, 545–577. [Google Scholar] [CrossRef]
- Khan, S.M.; Sung, J.Y.; Hebert, T.E. Gβγ subunits-Different spaces, different faces. Pharmacol. Res. 2016, 111, 434–441. [Google Scholar] [CrossRef]
- Lohmann, K.; Masuho, I.; Patil, D.N.; Baumann, H.; Hebert, E.; Steinrucke, S.; Trujillano, D.; Skamangas, N.K.; Dobricic, V.; Huning, I.; et al. Novel GNB1 mutations disrupt assembly and function of G protein heterotrimers and cause global developmental delay in humans. Hum. Mol. Genet. 2017, 26, 1078–1086. [Google Scholar] [CrossRef]
- Reddy, H.P.; Yakubovich, D.; Keren-Raifman, T.; Tabak, G.; Tsemakhovich, V.A.; Pedersen, M.H.; Shalomov, B.; Colombo, S.; Goldstein, D.B.; Javitch, J.A.; et al. Encephalopathy-causing mutations in Gβ(1) (GNB1) alter regulation of neuronal GIRK channels. iScience 2021, 24, 103018. [Google Scholar] [CrossRef]
- Steinrucke, S.; Lohmann, K.; Domingo, A.; Rolfs, A.; Baumer, T.; Spiegler, J.; Hartmann, C.; Munchau, A. Novel GNB1 missense mutation in a patient with generalized dystonia, hypotonia, and intellectual disability. Neurol. Genet. 2016, 2, e106. [Google Scholar] [CrossRef]
- Szczaluba, K.; Biernacka, A.; Szymanska, K.; Gasperowicz, P.; Kosinska, J.; Rydzanicz, M.; Ploski, R. Novel GNB1 de novo mutation in a patient with neurodevelopmental disorder and cutaneous mastocytosis: Clinical report and literature review. Eur. J. Med. Genet. 2018, 61, 157–160. [Google Scholar] [CrossRef] [PubMed]
- Okae, H.; Iwakura, Y. Neural tube defects and impaired neural progenitor cell proliferation in Gβ1-deficient mice. Dev. Dyn. 2010, 239, 1089–1101. [Google Scholar] [CrossRef] [PubMed]
- Dupre, D.J.; Baragli, A.; Rebois, R.V.; Ethier, N.; Hebert, T.E. Signalling complexes associated with adenylyl cyclase II are assembled during their biosynthesis. Cell. Signal. 2007, 19, 481–489. [Google Scholar] [CrossRef]
- Rebois, R.V.; Robitaille, M.; Petrin, D.; Zylbergold, P.; Trieu, P.; Hebert, T.E. Combining protein complementation assays with resonance energy transfer to detect multipartner protein complexes in living cells. Methods 2008, 45, 214–218. [Google Scholar] [CrossRef] [PubMed]
- David, M.; Richer, M.; Mamarbachi, A.M.; Villeneuve, L.R.; Dupre, D.J.; Hebert, T.E. Interactions between GABA-B1 receptors and Kir 3 inwardly rectifying potassium channels. Cell. Signal. 2006, 18, 2172–2181. [Google Scholar] [CrossRef]
- Liao, Y.; Sloan, D.C.; Widjaja, J.H.; Muntean, B.S. KCTD5 Forms Hetero-Oligomeric Complexes with Various Members of the KCTD Protein Family. Int. J. Mol. Sci. 2023, 24, 14317. [Google Scholar] [CrossRef]
- Balasco, N.; Esposito, L.; Smaldone, G.; Salvatore, M.; Vitagliano, L. A Comprehensive Analysis of the Structural Recognition between KCTD Proteins and Cullin 3. Int. J. Mol. Sci. 2024, 25, 1881. [Google Scholar] [CrossRef]
- Esposito, L.; Balasco, N.; Smaldone, G.; Berisio, R.; Ruggiero, A.; Vitagliano, L. AlphaFold-Predicted Structures of KCTD Proteins Unravel Previously Undetected Relationships among the Members of the Family. Biomolecules 2021, 11, 1862. [Google Scholar] [CrossRef]
- Esposito, L.; Balasco, N.; Vitagliano, L. Alphafold Predictions Provide Insights into the Structural Features of the Functional Oligomers of All Members of the KCTD Family. Int. J. Mol. Sci. 2022, 23, 13346. [Google Scholar] [CrossRef]
- Jones, D.T.; Taylor, W.R.; Thornton, J.M. The rapid generation of mutation data matrices from protein sequences. Comput. Appl. Biosci. 1992, 8, 275–282. [Google Scholar] [CrossRef]
- Tamura, K.; Stecher, G.; Kumar, S. MEGA11: Molecular Evolutionary Genetics Analysis Version 11. Mol. Biol. Evol. 2021, 38, 3022–3027. [Google Scholar] [CrossRef]
- Lukasheva, V.; Devost, D.; Le Gouill, C.; Namkung, Y.; Martin, R.D.; Longpre, J.M.; Amraei, M.; Shinjo, Y.; Hogue, M.; Lagace, M.; et al. Signal profiling of the β1AR reveals coupling to novel signalling pathways and distinct phenotypic responses mediated by β1AR and β2AR. Sci. Rep. 2020, 10, 8779. [Google Scholar] [CrossRef]
- Martin, R.D.; Sun, Y.; MacKinnon, S.; Cuccia, L.; Page, V.; Hebert, T.E.; Tanny, J.C. Differential Activation of P-TEFb Complexes in the Development of Cardiomyocyte Hypertrophy following Activation of Distinct G Protein-Coupled Receptors. Mol. Cell Biol. 2020, 40, e00048-20. [Google Scholar] [CrossRef]
- Jones-Tabah, J.; Martin, R.D.; Chen, J.J.; Tanny, J.C.; Clarke, P.B.S.; Hebert, T.E. A role for BET proteins in regulating basal, dopamine-induced and cAMP/PKA-dependent transcription in rat striatal neurons. Cell. Signal. 2022, 91, 110226. [Google Scholar] [CrossRef]
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Rizk, R.; Devost, D.; Pétrin, D.; Hébert, T.E. KCTD Proteins Have Redundant Functions in Controlling Cellular Growth. Int. J. Mol. Sci. 2024, 25, 4993. https://doi.org/10.3390/ijms25094993
Rizk R, Devost D, Pétrin D, Hébert TE. KCTD Proteins Have Redundant Functions in Controlling Cellular Growth. International Journal of Molecular Sciences. 2024; 25(9):4993. https://doi.org/10.3390/ijms25094993
Chicago/Turabian StyleRizk, Robert, Dominic Devost, Darlaine Pétrin, and Terence E. Hébert. 2024. "KCTD Proteins Have Redundant Functions in Controlling Cellular Growth" International Journal of Molecular Sciences 25, no. 9: 4993. https://doi.org/10.3390/ijms25094993