Affiliation:
1. Instituto de Bioquímica Vegetal y Fotosíntesis, Consejo Superior de Investigaciones Científicas and Universidad de Sevilla, Seville, Spain
Abstract
ABSTRACT
Heterocyst differentiation is orchestrated by the N control transcriptional regulator NtcA and the differentiation-specific factor HetR. In
Anabaena
sp. strain PCC 7120, the
devBCA
operon is expressed from two different promoters activated upon N stepdown. The distal
devB
promoter (transcription start point [TSP] located at position −704) represents a canonical class II NtcA-activated promoter, including a consensus NtcA-binding site centered 39.5 nucleotides upstream from the TSP. Transcription activation from a second TSP (−454) requires NtcA and is impaired in
hetR
mutants. In a wild-type background, three different DNA fragments, including both or each individual promoter, directed
gfp
expression localized mainly to proheterocysts and heterocysts. Expression was undetectable in an
ntcA
background and, for the fragment including the proximal promoter alone, also in a
hetR
background. In spite of the absence of consensus NtcA-binding sequences between the two TSPs, NtcA was shown to interact with this DNA region, and NtcA and its effector, 2-oxoglutarate, were necessary and sufficient for
in vitro
transcription from the −454 TSP. No HetR binding to the DNA or
in vitro
transcription from the proximal
devB
TSP promoted by HetR alone were detected. However, a moderate positive effect of HetR on NtcA binding to the DNA between the two
devB
TSPs was observed. The proximal
devB
promoter appears to represent a suboptimal NtcA-activated promoter for which HetR may act as a coactivator, with the physiological effect of restricting gene activation to conditions of prevalence of high NtcA and HetR levels, such as those taking place during heterocyst differentiation.
Publisher
American Society for Microbiology
Subject
Molecular Biology,Microbiology
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