Cloning of the mismatch recognition protein MSH2 from zebrafish ( Danio rerio) and its developmental stage-dependent mRNA expression

Abstract

Eukaryotic mismatch repair of simple base mispairs and small insertion-deletion loops is activated by the binding of a heterodimeric complex composed of MutS homolog 2(MSH2) and MSH6. Here we report the cloning of zebrafish ( Danio rerio) MSH2 (zMSH2) cDNA that has an open reading frame of 2811 nucleotides encoding a polypeptide of 936 amino acids. The deduced amino acid sequence of zMSH2 shares a 69% identity to both human and mouse MSH2. The zMSH2 protein contains a putative tyrosine-42 mismatch-contacting residue located at the N-terminal mismatch recognition region and four C-terminal ATP-binding consensus sequences conserved among MutS homologs. The 105-kDa recombinant zMSH2 bound apparently stronger to a G–T heteroduplex than to a homoduplex probe as shown by a gel shift assay. A preferential expression of both zMSH2 and zMSH6 mRNA in early embryos was found by Northern blot analysis. Whole mount in situ hybridization revealed a major expression of zMSH2 in different regions of the brain, including eyes, telencephalon, and the fourth ventricle in 12- to 48-h-old embryos. The production of zMSH2 mRNA gradually decreased in more mature 60- to 120-h-old zebrafish, reflecting a positive correlation between the amount of proliferating cells and MSH gene expression.