*Article* **Comparative Analysis of Proteins Regulated during Cadmium Sulfide Quantum Dots Response in** *Arabidopsis thaliana* **Wild Type and Tolerant Mutants**

**Valentina Gallo 1, Andrea Zappettini 2, Marco Villani 2, Nelson Marmiroli 1,3 and Marta Marmiroli 1,\***


**Abstract:** In previous work, two independent *Arabidopsis thaliana* Ac/Ds transposon insertional mutant lines, *atnp01* and *atnp02,* were identified that showed a higher level of tolerance than the wild type (wt) line to cadmium sulfide quantum dots (CdS QDs). The tolerance response was characterized at physiological, genetic and transcriptomic levels. In this work, a comparative analysis was performed on protein extracts from plantlets of the two mutants and of wt, each treated with 80 mg L−<sup>1</sup> CdS QDs. A comparative protein analysis was performed by 2D-PAGE, and proteins were characterized by MALDI-TOF/TOF mass spectrometry. Of 250 proteins identified from all three lines, 98 showed significant changes in relative abundance between control and CdS QD-treated plantlets. The wt, *atnp01*, and *atnp02* control-treated pairs respectively showed 61, 31, and 31 proteins with differential expression. The two mutants had a different response to treatment in terms of type and quantity of up- and downregulated proteins. This difference became more striking when compared to wt. A network analysis of the proteins differentially expressed in *atnp01* and *atnp02* included several of those encoded by putative genes accommodating the transposons, which were responsible for regulation of some proteins identified in this study. These included nifu-like protein 3 (Nfu3), involved in chloroplast assembly, elongator complex 3 (Elo3), involved in transcriptional elongation, magnesium-chelate subunit-2 (Chli2), involved in chlorophyll biosynthesis, and protein phosphatase 2C (PP2C) which mediates abiotic stress response.

**Keywords:** proteomics; engineered nanomaterials; mutants; 2D-PAGE; stress response proteins; network analysis
