Characterization of the ECB binding complex responsible for the M/G(1)-specific transcription of CLN3 and SWI4.

Abstract:

:The transcription factor Mcm1 is regulated by adjacent binding of a variety of different factors regulating the expression of cell-type-specific, cell cycle-specific, and metabolic genes. In this work, we investigate a new class of Mcm1-regulated promoters that are cell cycle regulated and peak in late M-early G(1) phase of the cell cycle via a promoter element referred to as an early cell cycle box (ECB). Gel filtration experiments indicate that the ECB-specific DNA binding complex is over 200 kDa in size and includes Mcm1 and at least one additional protein. Using DNase I footprinting in vitro, we have observed protection of the ECB elements from the CLN3, SWI4, CDC6, and CDC47 promoters, which includes protection of the 16-bp palindrome to which Mcm1 dimers are known to bind as well as protection of extended flanking sequences. These flanking sequences influence the stability and the variety of complexes that form on the ECB elements, and base substitutions in the protected flank affect transcriptional activity of the element. Chromatin immunoprecipitations show that Mcm1 binds in vivo to ECB elements throughout the cell cycle and that binding is sensitive to carbon source changes.

journal_name

Mol Cell Biol

authors

Mai B,Miles S,Breeden LL

doi

10.1128/mcb.22.2.430-441.2002

subject

Has Abstract

pub_date

2002-01-01 00:00:00

pages

430-41

issue

2

eissn

0270-7306

issn

1098-5549

journal_volume

22

pub_type

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