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Yeast screens identify the RNA polymerase II CTD and SPT5 as relevant targets of BRCA1 interaction.

dc.contributor.author Bennett, CB
dc.contributor.author Westmoreland, TJ
dc.contributor.author Verrier, CS
dc.contributor.author Blanchette, CA
dc.contributor.author Sabin, TL
dc.contributor.author Phatnani, HP
dc.contributor.author Mishina, YV
dc.contributor.author Huper, G
dc.contributor.author Selim, AL
dc.contributor.author Madison, ER
dc.contributor.author Bailey, DD
dc.contributor.author Falae, AI
dc.contributor.author Galli, A
dc.contributor.author Olson, JA
dc.contributor.author Greenleaf, AL
dc.contributor.author Marks, JR
dc.coverage.spatial United States
dc.date.accessioned 2011-06-21T17:31:22Z
dc.date.issued 2008-01-16
dc.identifier http://www.ncbi.nlm.nih.gov/pubmed/18197258
dc.identifier.uri http://hdl.handle.net/10161/4482
dc.description.abstract BRCA1 has been implicated in numerous DNA repair pathways that maintain genome integrity, however the function responsible for its tumor suppressor activity in breast cancer remains obscure. To identify the most highly conserved of the many BRCA1 functions, we screened the evolutionarily distant eukaryote Saccharomyces cerevisiae for mutants that suppressed the G1 checkpoint arrest and lethality induced following heterologous BRCA1 expression. A genome-wide screen in the diploid deletion collection combined with a screen of ionizing radiation sensitive gene deletions identified mutants that permit growth in the presence of BRCA1. These genes delineate a metabolic mRNA pathway that temporally links transcription elongation (SPT4, SPT5, CTK1, DEF1) to nucleopore-mediated mRNA export (ASM4, MLP1, MLP2, NUP2, NUP53, NUP120, NUP133, NUP170, NUP188, POM34) and cytoplasmic mRNA decay at P-bodies (CCR4, DHH1). Strikingly, BRCA1 interacted with the phosphorylated RNA polymerase II (RNAPII) carboxy terminal domain (P-CTD), phosphorylated in the pattern specified by the CTDK-I kinase, to induce DEF1-dependent cleavage and accumulation of a RNAPII fragment containing the P-CTD. Significantly, breast cancer associated BRCT domain defects in BRCA1 that suppressed P-CTD cleavage and lethality in yeast also suppressed the physical interaction of BRCA1 with human SPT5 in breast epithelial cells, thus confirming SPT5 as a relevant target of BRCA1 interaction. Furthermore, enhanced P-CTD cleavage was observed in both yeast and human breast cells following UV-irradiation indicating a conserved eukaryotic damage response. Moreover, P-CTD cleavage in breast epithelial cells was BRCA1-dependent since damage-induced P-CTD cleavage was only observed in the mutant BRCA1 cell line HCC1937 following ectopic expression of wild type BRCA1. Finally, BRCA1, SPT5 and hyperphosphorylated RPB1 form a complex that was rapidly degraded following MMS treatment in wild type but not BRCA1 mutant breast cells. These results extend the mechanistic links between BRCA1 and transcriptional consequences in response to DNA damage and suggest an important role for RNAPII P-CTD cleavage in BRCA1-mediated cancer suppression.
dc.language eng
dc.language.iso en_US
dc.relation.ispartof PLoS One
dc.relation.isversionof 10.1371/journal.pone.0001448
dc.subject BRCA1 Protein
dc.subject Cell Cycle
dc.subject Chromosomal Proteins, Non-Histone
dc.subject DNA Damage
dc.subject Genes, Lethal
dc.subject Genomic Instability
dc.subject Humans
dc.subject Hydrolysis
dc.subject RNA Polymerase II
dc.subject Transcriptional Elongation Factors
dc.title Yeast screens identify the RNA polymerase II CTD and SPT5 as relevant targets of BRCA1 interaction.
dc.title.alternative
dc.type Journal article
dc.description.version Version of Record
duke.date.pubdate 2008-1-16
duke.description.issue 1
duke.description.volume 3
dc.relation.journal Plos One
pubs.author-url http://www.ncbi.nlm.nih.gov/pubmed/18197258
pubs.begin-page e1448
pubs.issue 1
pubs.organisational-group Basic Science Departments
pubs.organisational-group Biochemistry
pubs.organisational-group Clinical Science Departments
pubs.organisational-group Duke
pubs.organisational-group Duke Cancer Institute
pubs.organisational-group Institutes and Centers
pubs.organisational-group Molecular Genetics and Microbiology
pubs.organisational-group Pathology
pubs.organisational-group School of Medicine
pubs.organisational-group Surgery
pubs.organisational-group Surgery, Surgical Sciences
pubs.publication-status Published online
pubs.volume 3
dc.identifier.eissn 1932-6203


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