Modular recognition of 5-base-pair DNA sequence motifs by human heat shock transcription factor.

Abstract:

:We investigated the recognition of the conserved 5-bp repeated motif NGAAN, which occurs in heat shock gene promoters of Drosophila melanogaster and other eukaryotic organisms, by human heat shock transcription factor (HSF). Extended heat shock element mutants of the human HSP70 gene promoter, containing additional NGAAN blocks flanking the original element, showed significantly higher affinity than the wild-type promoter element for human HSF in vitro. Protein-DNA contact positions were identified by hydroxyl radical protection, diethyl pyrocarbonate interference, and DNase I footprinting. New contacts in the mutant HSE constructs corresponded to the locations of additional NGAAN motifs. The pattern of binding indicated the occurrence of multiple DNA binding modes for HSF with the various constructs and was consistent with an oligomeric, possibly trimeric, structure of the protein. In contrast to the improved binding, the extended heat shock element mutant constructs did not exhibit dramatically increased heat-inducible transcription in transient expression assays with HeLa cells.

journal_name

Mol Cell Biol

authors

Cunniff NF,Wagner J,Morgan WD

doi

10.1128/mcb.11.7.3504

subject

Has Abstract

pub_date

1991-07-01 00:00:00

pages

3504-14

issue

7

eissn

0270-7306

issn

1098-5549

journal_volume

11

pub_type

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