Abstract
The yeast SNM1/PSO2 gene specifically functions in DNA interstrand cross-link (ICL) repair, and its role has been suggested to be separate from other DNA repair pathways. In vertebrates, there are three homologs of SNM1 (SNM1A, SNM1B, and SNM1C/Artemis; SNM1 family proteins) whose functions are largely unknown. We disrupted each of the SNM1 family genes in the chicken B-cell line DT40. Both SNM1A- and SNM1B-deficient cells were sensitive to cisplatin but not to X-rays, whereas SNM1C/Artemis-deficient cells exhibited sensitivity to X-rays but not to cisplatin. SNM1A was nonepistatic with XRCC3 (homologous recombination), RAD18 (translesion synthesis), FANCC (Fanconi anemia), and SNM1B in ICL repair. SNM1A protein formed punctate nuclear foci depending on the conserved SNM1 (metallo-beta-lactamase) domain. PIAS1 was found to physically interact with SNM1A, and they colocalized at nuclear foci. Point mutations in the SNM1 domain, which disrupted the interaction with PIAS1, led to mislocalization of SNM1A in the nucleus and loss of complementation of snm1a cells. These results suggest that interaction between SNM1A and PIAS1 is required for ICL repair.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Amino Acid Sequence
-
Animals
-
Cell Line
-
Cell Line, Tumor
-
Cell Nucleus / metabolism
-
Chickens
-
Cisplatin / pharmacology
-
Cross-Linking Reagents / pharmacology
-
DNA Mutational Analysis
-
DNA Repair
-
DNA Repair Enzymes
-
DNA, Fungal / genetics
-
DNA, Fungal / metabolism*
-
DNA-Binding Proteins / chemistry
-
DNA-Binding Proteins / genetics
-
DNA-Binding Proteins / metabolism*
-
Endodeoxyribonucleases
-
Exodeoxyribonucleases
-
Fluorescent Dyes
-
Fungal Proteins / chemistry
-
Fungal Proteins / genetics
-
Fungal Proteins / metabolism*
-
HeLa Cells
-
Humans
-
Microscopy, Fluorescence
-
Molecular Sequence Data
-
Nuclear Proteins / chemistry
-
Nuclear Proteins / genetics
-
Nuclear Proteins / metabolism*
-
Organic Chemicals
-
Point Mutation
-
Protein Inhibitors of Activated STAT
-
Protein Structure, Tertiary
-
Recombination, Genetic
-
Saccharomyces cerevisiae Proteins / chemistry
-
Saccharomyces cerevisiae Proteins / genetics
-
Saccharomyces cerevisiae Proteins / metabolism*
-
Small Ubiquitin-Related Modifier Proteins / metabolism*
-
Two-Hybrid System Techniques
-
X-Rays
Substances
-
Alexa594
-
Cross-Linking Reagents
-
DNA, Fungal
-
DNA-Binding Proteins
-
Fluorescent Dyes
-
Fungal Proteins
-
Nuclear Proteins
-
Organic Chemicals
-
PIAS1 protein, human
-
Protein Inhibitors of Activated STAT
-
Saccharomyces cerevisiae Proteins
-
Small Ubiquitin-Related Modifier Proteins
-
DCLRE1B protein, human
-
Endodeoxyribonucleases
-
Exodeoxyribonucleases
-
PSO2 protein, S cerevisiae
-
DNA Repair Enzymes
-
Cisplatin
Associated data
-
GENBANK/AY376896
-
GENBANK/AY376897
-
GENBANK/AY376898
-
GENBANK/AY549568