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PMID: 8978690 Published · ppublish English Comparative Study Journal Article

The Schizosaccharomyces pombe rad3 checkpoint gene.

The EMBO journal ·Vol. 15 ·No. 23 ·1996-12-02 ·Pages 6641-51

Bentley NJ, Holtzman DA, Flaggs G, Keegan KS, DeMaggio A, Ford JC, Hoekstra M, Carr AM

Abstract

The rad3 gene of Schizosaccharomyces pombe is required for checkpoint pathways that respond to DNA damage and replication blocks. We report the complete rad3 gene sequence and show that rad3 is the homologue of Saccharomyces cerevisiae ESR1 (MEC1/SAD3) and Drosophila melanogaster mei-41 checkpoint genes. This establishes Rad3/Mec1 as the only conserved protein which is required for all the DNA structure checkpoints in both yeast model systems. Rad3 is an inessential member of the 'lipid kinase' subclass of kinases which includes the ATM protein defective in ataxia telangiectasia patients. Mutational analysis indicates that the kinase domain is required for Rad3 function, and immunoprecipitation of overexpressed Rad3 demonstrates an associated protein kinase activity. The previous observation that rad3 mutations can be rescued by a truncated clone lacking the kinase domain may be due to intragenic complementation. Consistent with this, biochemical data suggest that Rad3 exists in a complex containing multiple copies of Rad3. We have identified a novel human gene (ATR) whose product is closely related to Rad3/Esr1p/Mei-41. ATR can functionally complement esr1-1 radiation sensitivity in S. cerevisiae. Together, the structural conservation and functional complementation suggest strongly that the mechanisms underlying the DNA structure checkpoints are conserved throughout evolution.

MeSH Terms
Adenosine Triphosphatases/chemistry,genetics,metabolism Amino Acid Sequence Animals Ataxia Telangiectasia Mutated Proteins Base Sequence Cell Cycle Proteins/chemistry,metabolism Cloning, Molecular DNA Damage DNA Helicases/chemistry,genetics,metabolism DNA Primers DNA Replication Dose-Response Relationship, Radiation Drosophila melanogaster/genetics Genes, Fungal Genetic Complementation Test Humans Kinetics Molecular Sequence Data Mutagenesis Oligodeoxyribonucleotides Phosphotransferases/metabolism Phylogeny Protein Serine-Threonine Kinases Recombinant Proteins/chemistry,metabolism Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins Schizosaccharomyces/cytology,genetics,radiation effects Sequence Deletion Sequence Homology, Amino Acid
Chemicals
Cell Cycle Proteins DNA Primers Oligodeoxyribonucleotides Recombinant Proteins Saccharomyces cerevisiae Proteins Phosphotransferases ATR protein, human Ataxia Telangiectasia Mutated Proteins Protein Serine-Threonine Kinases Adenosine Triphosphatases Rad3 protein, S cerevisiae DNA Helicases
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Bentley N J
MRC Cell Mutation Unit, Sussex University, Falmer, UK.
Holtzman D A
Flaggs G
Keegan K S
DeMaggio A
Ford J C
Hoekstra M
Carr A M
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1996-12-02
Pages
6641-51
Language
English
Region
England
NLM ID
8208664
PMCID
PMC452488
Subset
IM
Databases
GENBANK
U76307, U76308, Y09076, Y09077
External Links
PubMed source PMC Full Text View journal details
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