A portable BRCA1-HAC (human artificial chromosome) module for analysis of BRCA1 tumor suppressor function

dc.contributor.authorKononenko, Artem V.
dc.contributor.authorBansal, Ruchi
dc.contributor.authorLee, Nicholas C.O.
dc.contributor.authorGrimes, Brenda R.
dc.contributor.authorMasumoto, Hiroshi
dc.contributor.authorEarnshaw, William C.
dc.contributor.authorLarionov, Vladimir
dc.contributor.authorKouprina, Natalay
dc.contributor.departmentDepartment of Medical & Molecular Genetics, IU School of Medicineen_US
dc.date.accessioned2016-03-18T15:06:15Z
dc.date.available2016-03-18T15:06:15Z
dc.date.issued2014-12-01
dc.description.abstractBRCA1 is involved in many disparate cellular functions, including DNA damage repair, cell-cycle checkpoint activation, gene transcriptional regulation, DNA replication, centrosome function and others. The majority of evidence strongly favors the maintenance of genomic integrity as a principal tumor suppressor activity of BRCA1. At the same time some functional aspects of BRCA1 are not fully understood. Here, a HAC (human artificial chromosome) module with a regulated centromere was constructed for delivery and expression of the 90 kb genomic copy of the BRCA1 gene into BRCA1-deficient human cells. A battery of functional tests was carried out to demonstrate functionality of the exogenous BRCA1. In separate experiments, we investigated the role of BRCA1 in maintenance of heterochromatin integrity within a human functional kinetochore. We demonstrated that BRCA1 deficiency results in a specific activation of transcription of higher-order alpha-satellite repeats (HORs) assembled into heterochromatin domains flanking the kinetochore. At the same time no detectable elevation of transcription was observed within HORs assembled into centrochromatin domains. Thus, we demonstrated a link between BRCA1 deficiency and kinetochore dysfunction and extended previous observations that BRCA1 is required to silence transcription in heterochromatin in specific genomic loci. This supports the hypothesis that epigenetic alterations of the kinetochore initiated in the absence of BRCA1 may contribute to cellular transformation.en_US
dc.eprint.versionFinal published versionen_US
dc.identifier.citationKononenko, A. V., Bansal, R., Lee, N. C. O., Grimes, B. R., Masumoto, H., Earnshaw, W. C., … Kouprina, N. (2014). A portable BRCA1-HAC (human artificial chromosome) module for analysis of BRCA1 tumor suppressor function. Nucleic Acids Research, 42(21), e164. http://doi.org/10.1093/nar/gku870en_US
dc.identifier.issn0305-1048en_US
dc.identifier.urihttps://hdl.handle.net/1805/8925
dc.language.isoen_USen_US
dc.publisherOxford University Pressen_US
dc.relation.isversionof10.1093/nar/gku870en_US
dc.relation.journalNucleic Acids Researchen_US
dc.rightsCC0 1.0 Universal en_US
dc.rights.urihttps://creativecommons.org/publicdomain/zero/1.0
dc.sourcePublisheren_US
dc.subjectBRCA1 Proteinen_US
dc.subjectmetabolismen_US
dc.subjectChromosomes, Artificial, Humanen_US
dc.subjectGenes, BRCA1en_US
dc.titleA portable BRCA1-HAC (human artificial chromosome) module for analysis of BRCA1 tumor suppressor functionen_US
dc.typeArticleen_US
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