Discovery, expression, cellular localization, and molecular properties of a novel, alternative spliced HP1γ isoform, lacking the chromoshadow domain

• Published in: PloS one 2020-02, Vol.15 (2), p.e0217452
• Main Authors: Mathison, Angela, Milech De Assuncao, Thiago, Dsouza, Nikita R, Williams, Monique, Zimmermann, Michael T, Urrutia, Raul, Lomberk, Gwen
• Format: Article
• Language: English
• Subjects: Alternative Splicing; Animals; Binding Sites; Bioinformatics; Biology and life sciences; Cancer; Cell differentiation; Cell proliferation; CHO Cells; Chromobox Protein Homolog 5; Chromosomal Proteins, Non-Histone - chemistry; Chromosomal Proteins, Non-Histone - genetics; Chromosomal Proteins, Non-Histone - metabolism; Computer simulation; Cricetinae; Cricetulus; Deoxyribonucleic acid; Dimerization; DNA; DNA repair; Domains; Gene Expression Regulation, Neoplastic; Genomics; HeLa Cells; Histones - metabolism; Homology; Human tissues; Humans; Insulin-Secreting Cells - metabolism; Isoforms; Laboratories; Localization; Medical schools; Molecular dynamics; Molecular modelling; Molecular properties; mRNA; Physical Sciences; Precision medicine; Protein Binding; Protein folding; Protein Transport; Proteins; R&D; Research & development; Research and Analysis Methods; Surgery
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0217452

By reading the H3K9Me3 mark through their N-terminal chromodomain (CD), HP1 proteins play a significant role in cancer-associated processes, including cell proliferation, differentiation, chromosomal stability, and DNA repair. Here, we used a combination of bioinformatics-based methodologies, as well as experimentally-derived datasets, that reveal the existence of a novel short HP1γ (CBX3) isoform, named here sHP1γ, generated by alternative splicing of the CBX3 locus. The sHP1γ mRNA encodes a protein composed of 101 residues and lacks the C-terminal chromoshadow domain (CSD) that is required for dimerization and heterodimerization in the previously described 183 a. a HP1γ protein. Fold recognition, order-to-disorder calculations, threading, homology-based molecular modeling, docking, and molecular dynamic simulations show that the sHP1γ is comprised of a CD flanked by intrinsically disordered regions (IDRs) with an IDR-CD-IDR domain organization and likely retains the ability to bind to the H3K9Me3. Both qPCR analyses and mRNA-seq data derived from large-scale studies confirmed that sHP1γ mRNA is expressed in the majority of human tissues at approximately constant ratios with the chromoshadow domain containing isoform. However, sHP1γ mRNA levels appear to be dysregulated in different cancer types. Thus, our data supports the notion that, due to the existence of functionally different isoforms, the regulation of HP1γ-mediated functions is more complex than previously anticipated.