The high mobility group domain protein Cmb1 of Schizosaccharomyces pombe binds to cytosines in base mismatches and opposite chemically altered guanines.
The mismatch-binding activity Cmb1 of Schizosaccharomyces pombe was enriched from wild type cells, and N-terminal sequencing enabled cloning of the respective gene. The deduced amino acid sequence of cmb1(+) contains a high mobility group domain, a motif that is common to a heterogeneous family of DNA-binding proteins. In crude protein extracts of a cmb1 gene-disruption strain, specific binding to C/T, C/A, and C/Delta was abolished. Weak binding to C/C revealed the presence of a second mismatch-binding activity, Cmb2. Cmb1, enriched from S. pombe and purified from Escherichia coli, bound specifically to C/C, C/T, C/A, T/T, and C/Delta but showed little or no affinity to other mismatches and small loops. Cmb1 recognizes 1,2 GpG intrastrand cross-links, produced by the chemotherapeutic drug cisplatin, when two cytosines are opposite the cross-linked guanines but not when other bases are present. Consistently, O6-methylguanine:C but not O6-methylguanine/T lesions were bound. Thus, cytosines in mismatches and opposite chemically modified guanines are the preferred target of Cmb1 recognition. cmb1 mutant cells are more sensitive to cisplatin than wild type cells, indicating a role of Cmb1 in repair of cisplatin-induced DNA damage.
Fleck
O
O
Institute of General Microbiology, University of Bern, Baltzer-Strasse 4, CH-3012 Bern, Switzerland. fleck@imb.uibe.ch
Kunz
C
C
Rudolph
C
C
Kohli
J
J
D000595:Amino Acid Sequence; D020137:Base Pair Mismatch; D001483:Base Sequence; D002945:Cisplatin / Q000494:pharmacology; D003001:Cloning, Molecular; D003432:Cross-Linking Reagents / Q000494:pharmacology; D003596:Cytosine / Q000378:metabolism*; D004271:DNA, Fungal / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D006147:Guanine / Q000378:metabolism*; D006609:High Mobility Group Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology; D008969:Molecular Sequence Data; D012568:Schizosaccharomyces / Q000187:drug effects / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins
C115714:Cmb1 protein, S pombe; D003432:Cross-Linking Reagents; D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D029702:Schizosaccharomyces pombe Proteins; D008769:Methylnitronitrosoguanidine; C034697:transplatin; D006147:Guanine; D003596:Cytosine; D002945:Cisplatin