Exploring the roles of different DNA Repair Proteins in Short Inverted Repeat Mediated Genomic Instability: A Pilot...

Repetitive DNA sequences are abundant in the human genome and can adopt alternative (i.e., non-B) DNA structures. These sequences contribute to diverse biological functions, including genomic instability. Previously, we found that Z-DNA-, H-DNA-, and cruciform DNA-forming sequences are mutagenic, implicating them in cancer etiology. These sequences can stimulate the formation of DNA double-strand breaks (DSBs), causing deletions via cleavage by the endonuclease ERCC1-XPF. Interestingly, the activity of ERCC1-XPF in H-DNA-induced mutagenesis is nucleotide excision repair (NER)-dependent, but its role in Z-DNA-induced mutagenesis is NER-independent. Instead, Z-DNA is processed by ERCC1-XPF in a mechanism dependent on the mismatch repair (MMR) complex, MSH2-MSH3. These observations indicate distinct mechanisms of non-B-induced genomic instability. However, the roles of NER and MMR proteins and additional nucleases (CtIP and MRE11) in the processing of cruciform DNA remain unknown. Here, we present data on the processing of cruciform-forming short-perfect inverted repeats (IRs) by DNA repair proteins using mammalian cell-based systems. From this pilot study, we show that in contrast to H-DNA and Z-DNA, short perfect IRs are processed in a NER- and MMR-independent manner, and the nucleases CtIP and MRE11 suppress short perfect IR-induced genomic instability in mammalian cells.