(Phospho)proteomic Profiling of Microsatellite Unstable CRC Cells Reveals Alterations in Nuclear Signaling and Cholesterol Metabolism Caused by Frameshift Mutation of NMD Regulator UPF3A.

Abstract

DNA mismatch repair-deficient colorectal cancers (CRCs) accumulate numerous frameshift mutations at repetitive sequences recognized as microsatellite instability (MSI). When coding mononucleotide repeats (cMNRs) are affected, tumors accumulate frameshift mutations and premature termination codons (PTC) potentially leading to truncated proteins. Nonsense-mediated RNA decay (NMD) can degrade PTC-containing transcripts and protect from such faulty proteins. As it also regulates normal transcripts and cellular physiology, we tested whether NMD genes themselves are targets of MSI frameshift mutations. A high frequency of cMNR frameshift mutations in the UPF3A gene was found in MSI CRC cell lines (67.7%), MSI colorectal adenomas (55%) and carcinomas (63%). In normal colonic crypts, UPF3A expression was restricted to single chromogranin A-positive cells. SILAC-based proteomic analysis of KM12 CRC cells revealed UPF3A-dependent down-regulation of several enzymes involved in cholesterol biosynthesis. Furthermore, reconstituted UPF3A expression caused alterations of 85 phosphosites in 52 phosphoproteins. Most of them (38/52, 73%) reside in nuclear phosphoproteins involved in regulation of gene expression and RNA splicing. Since UPF3A mutations can modulate the (phospho)proteomic signature and expression of enzymes involved in cholesterol metabolism in CRC cells, UPF3A may influence other processes than NMD and loss of UPF3A expression might provide a growth advantage to MSI CRC cells.

• Authors: Malwina Michalak, Eva-Maria Katzenmaier, Nina Roeckel, Stefan M Woerner, Vera Fuchs, Uwe Warnken, Yan P Yuan, Peer Bork, Gabriele Neu-Yilik, Andreas Kulozik, Magnus von Knebel Doeberitz, Matthias Kloor, Jürgen Kopitz, Johannes Gebert
• Journal: International journal of molecular sciences (Int J Mol Sci) Vol. 21 Issue 15 (Jul 23 2020) ISSN: 1422-0067 [Electronic] Switzerland
• PMID: 32718059 (Publication Type: Journal Article)

Chemical References

• Neoplasm Proteins
• Phosphoproteins
• RNA-Binding Proteins
• UPF3A protein, human

Topics

• Cell Line, Tumor
• Colorectal Neoplasms (genetics, metabolism, pathology)
• Frameshift Mutation
• Gene Expression Profiling
• Genomic Instability
• Humans
• Microsatellite Repeats
• Neoplasm Proteins (genetics, metabolism)
• Nonsense Mediated mRNA Decay
• Phosphoproteins (genetics, metabolism)
• Proteomics
• RNA-Binding Proteins (genetics, metabolism)