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PMID: 14560028 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Mutations in yeast replication proteins that increase CAG/CTG expansions also increase repeat fragility.

Molecular and cellular biology ·Vol. 23 ·No. 21 ·2003-11-00 ·Pages 7849-60

Callahan JL, Andrews KJ, Zakian VA, Freudenreich CH

Abstract

Expansion of trinucleotide repeats (TNRs) is the causative mutation in several human genetic diseases. Expanded TNR tracts are both unstable (changing in length) and fragile (displaying an increased propensity to break). We have investigated the relationship between fidelity of lagging-strand replication and both stability and fragility of TNRs. We devised a new yeast artificial chromomosme (YAC)-based assay for chromosome breakage to analyze fragility of CAG/CTG tracts in mutants deficient for proteins involved in lagging-strand replication: Fen1/Rad27, an endo/exonuclease involved in Okazaki fragment maturation, the nuclease/helicase Dna2, RNase HI, DNA ligase, polymerase delta, and primase. We found that deletion of RAD27 caused a large increase in breakage of short and long CAG/CTG tracts, and defects in DNA ligase and primase increased breakage of long tracts. We also found a correlation between mutations that increase CAG/CTG tract breakage and those that increase repeat expansion. These results suggest that processes that generate strand breaks, such as faulty Okazaki fragment processing or DNA repair, are an important source of TNR expansions.

MeSH Terms
Adenosine Triphosphatases/genetics,metabolism Biological Assay Chromosomes, Artificial DNA Damage DNA Helicases/genetics,metabolism DNA Ligases/genetics,metabolism DNA Polymerase III/genetics,metabolism DNA Primase/genetics,metabolism DNA Replication Flap Endonucleases/genetics,metabolism Fungal Proteins/genetics,metabolism Humans Mutation Reproducibility of Results Ribonuclease H/genetics,metabolism Saccharomyces cerevisiae Proteins Trinucleotide Repeat Expansion Yeasts/genetics,metabolism
Chemicals
Fungal Proteins Saccharomyces cerevisiae Proteins DNA Primase DNA Polymerase III Flap Endonucleases Ribonuclease H ribonuclease HI Adenosine Triphosphatases DNA Helicases DNA2 protein, S cerevisiae DNA Ligases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Callahan Julie L
Department of Biology, Program in Genetics, Tufts University, Medford, Massachusetts 02155, USA.
Andrews Kenneth J
Zakian Virginia A
Freudenreich Catherine H
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.23.21.7849-7860.2003
Published
2003-11-00
Pages
7849-60
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC207578
Subset
IM
Grants
NCI NIH HHS · R01 CA079441 · United States
NIGMS NIH HHS · R01 GM063066 · United States
NCI NIH HHS · CA79441 · United States
NIGMS NIH HHS · GM63066 · United States
External Links
PubMed source DOI: 10.1128/MCB.23.21.7849-7860.2003 PMC Full Text View journal details
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