UV-induced hyperphosphorylation of replication protein a depends on DNA replication and expression of ATM protein

Gregory G Oakley, L. I. Loberg, J. Yao, M. A. Risinger, R. L. Yunker, M. Zernik-Kobak, K. K. Khanna, M. F. Lavin, M. P. Carty, K. Dixon

Research output: Contribution to journalArticle

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Abstract

Exposure to DNA-damaging agents triggers signal transduction pathways that are thought to play a role in maintenance of genomic stability. A key protein in the cellular processes of nucleotide excision repair, DNA recombination, and DNA double-strand break repair is the single-stranded DNA binding protein, RPA. We showed previously that the p34 subunit of RPA becomes hyperphosphorylated as a delayed response (4-8 h) to UV radiation (10-30 j/m2). Here we show that UV-induced RPA-p34 hyperphosphorylation depends on expression of ATM, the product of the gene mutated in the human genetic disorder ataxia telangiectasia (A-T). UV-induced RPA-p34 hyperphosphorylation was not observed in A-T cells, but this response was restored by ATM expression. Furthermore, purified ATM kinase phosphorylates the p34 subunit of RPA complex in vitro at many of the same sites that are phosphorylated in vivo after UV radiation. Induction of this DNA damage response was also dependent on DNA replication; inhibition of DNA replication by aphidicolin prevented induction of RPA-p34 hyperphosphorylation by UV radiation. We postulate that this pathway is triggered by the accumulation of aberrant DNA replication intermediates, resulting from DNA replication fork blockage by UV photoproducts. Further, we suggest that RPA-p34 is hyperphosphorylated as a participant in the recombinational postreplication repair of these replication products. Successful resolution of these replication intermediates reduces the accumulation of chromosomal aberrations that would otherwise occur as a consequence of UV radiation.

Original languageEnglish (US)
Pages (from-to)1199-1213
Number of pages15
JournalMolecular biology of the cell
Volume12
Issue number5
DOIs
StatePublished - Jan 1 2001

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Ataxia Telangiectasia Mutated Proteins
DNA Replication
Radiation
Ataxia Telangiectasia
Proteins
Aphidicolin
Inborn Genetic Diseases
Double-Stranded DNA Breaks
Genomic Instability
DNA
Medical Genetics
DNA-Binding Proteins
Chromosome Aberrations
DNA Repair
Genetic Recombination
DNA Damage
Signal Transduction
Phosphotransferases
Maintenance
Genes

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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UV-induced hyperphosphorylation of replication protein a depends on DNA replication and expression of ATM protein. / Oakley, Gregory G; Loberg, L. I.; Yao, J.; Risinger, M. A.; Yunker, R. L.; Zernik-Kobak, M.; Khanna, K. K.; Lavin, M. F.; Carty, M. P.; Dixon, K.

In: Molecular biology of the cell, Vol. 12, No. 5, 01.01.2001, p. 1199-1213.

Research output: Contribution to journalArticle

Oakley, GG, Loberg, LI, Yao, J, Risinger, MA, Yunker, RL, Zernik-Kobak, M, Khanna, KK, Lavin, MF, Carty, MP & Dixon, K 2001, 'UV-induced hyperphosphorylation of replication protein a depends on DNA replication and expression of ATM protein', Molecular biology of the cell, vol. 12, no. 5, pp. 1199-1213. https://doi.org/10.1091/mbc.12.5.1199
Oakley, Gregory G ; Loberg, L. I. ; Yao, J. ; Risinger, M. A. ; Yunker, R. L. ; Zernik-Kobak, M. ; Khanna, K. K. ; Lavin, M. F. ; Carty, M. P. ; Dixon, K. / UV-induced hyperphosphorylation of replication protein a depends on DNA replication and expression of ATM protein. In: Molecular biology of the cell. 2001 ; Vol. 12, No. 5. pp. 1199-1213.
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