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PMID: 8552078 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Distinct cis-acting elements mediate clock, light, and developmental regulation of the Neurospora crassa eas (ccg-2) gene.

Molecular and cellular biology ·Vol. 16 ·No. 2 ·1996-02-00 ·Pages 513-21

Bell-Pedersen D, Dunlap JC, Loros JJ

Abstract

The Neurospora crassa eas (ccg-2) gene, which encodes a fungal hydrophobin, is transcriptionally regulated by the circadian clock. In addition, eas (ccg-2) is positively regulated by light and transcripts accumulate during asexual development. To sort out the basis of this complex regulation, deletion analyses of the eas (ccg-2) promoter were carried out to localize the cis-acting elements mediating clock, light, and developmental control. The primary sequence determinants of a positive activating clock element (ACE) were found to reside in a 45-bp region, just upstream from the TATA box. Using a novel unregulated promoter/reporter system developed for this study, we show that a 68-bp sequence encompassing the ACE is sufficient to confer clock regulation on the eas (ccg-2) gene. Electrophoretic mobility shift assays using the ACE reveal factors present in N. crassa protein extracts that recognize and bind specifically to DNA containing this element. Separate regions of the eas (ccg-2) promoter involved in light induction and developmental control are identified and shown not to be required for clock-regulated expression of eas (ccg-2). The distinct nature of the ACE validates its use as a tool for the identification of upstream regulatory factors involved in clock control of gene expression.

MeSH Terms
Base Sequence Biological Clocks/genetics Circadian Rhythm/genetics DNA Mutational Analysis Fungal Proteins/genetics Gene Expression Regulation, Fungal Light Molecular Probe Techniques Molecular Sequence Data Neurospora crassa/genetics,radiation effects Promoter Regions, Genetic Protein Binding Sequence Deletion Transcription, Genetic
Chemicals
Fungal Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Bell-Pedersen D
Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755, USA.
Dunlap J C
Loros J J
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.16.2.513
Published
1996-02-00
Pages
513-21
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC231029
Subset
IM
Grants
NIGMS NIH HHS · GM15185 · United States
NIMH NIH HHS · MH01186 · United States
NIMH NIH HHS · MH44651 · United States
External Links
PubMed source DOI: 10.1128/MCB.16.2.513 PMC Full Text View journal details
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