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The specification of mammalian chromosomal replication origins is incompletely understood. To analyze the assembly and activation of prereplicative complexes (pre-RCs), we tested the effects of tethered binding of chromatin acetyltransferases and replication proteins on chromosomal c-myc origin deletion mutants containing a GAL4-binding cassette. GAL4DBD (DNA binding domain) fusions with Orc2, Cdt1, E2F1 or HBO1 coordinated the recruitment of the Mcm7 helicase subunit, the DNA unwinding element (DUE)-binding protein DUE-B and the minichromosome maintenance (MCM) helicase activator Cdc45 to the replicator, and restored origin activity. In contrast, replication protein binding and origin activity were not stimulated by fusion protein binding in the absence of flanking c-myc DNA. Substitution of the GAL4-binding site for the c-myc replicator DUE allowed Orc2 and Mcm7 binding, but eliminated origin activity, indicating that the DUE is essential for pre-RC activation. Additionally, tethering of DUE-B was not sufficient to recruit Cdc45 or activate pre-RCs formed in the absence of a DUE. These results show directly in a chromosomal background that chromatin acetylation, Orc2 or Cdt1 suffice to recruit all downstream replication initiation activities to a prospective origin, and that chromosomal origin activity requires singular DNA sequences. 相似文献
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Grzeskowiak R Amin J Oetjen E Knepel W 《The Journal of biological chemistry》2000,275(39):30037-30045
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DNA binding and antigene activity of a daunomycin-conjugated triplex-forming oligonucleotide targeting the P2 promoter of the human c-myc gene 总被引:2,自引:2,他引:0
Carbone GM McGuffie E Napoli S Flanagan CE Dembech C Negri U Arcamone F Capobianco ML Catapano CV 《Nucleic acids research》2004,32(8):2396-2410
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TGF-beta regulation of epithelial cell proliferation. 总被引:4,自引:0,他引:4
H L Moses 《Molecular reproduction and development》1992,32(2):179-184
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Spedale G Meddens CA Koster MJ Ko CW van Hooff SR Holstege FC Timmers HT Pijnappel WW 《Nucleic acids research》2012,40(3):996-1008
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Hsien-Ming Hu Magdalena Arcinas Linda M Boxer 《The Journal of biological chemistry》2002,277(12):9819-9824
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