Characterization and transcriptional analysis of hupSLW in Gloeothece sp ATCC 27152: an uptake hydrogenase from a unicellular cyanobacterium

• Type: Article in International Scientific Journal
• Date: 2004

Authors

Oliveira, P (Author) Other; Leitao, E (Author) Other; Tamagnini, P (Author) FCUP; Moradas Ferreira, P (Author) ICBAS; Oxelfelt, F (Author) Other

Journal

Title: Microbiology

Vol. 150

Pages: 3647-3655

ISSN: 1350-0872

Publisher: Microbiological Society

Indexing

ISI Web of Knowledge - 32 Citations

Scopus - 32 Citations

Scientific classification

FOS: Natural sciences > Biological sciences

Other information

Authenticus ID: P-000-7ZB

DOI: 10.1099/mic.0.27248-0

Abstract (EN): The structural genes (hupSL) encoding an uptake hydrogenase in the unicellular cyanobacterium Gloeothece sp. ATCC 27152, a strain capable of aerobic N-2 fixation, were identified and sequenced, 3'-RACE experiments uncovered the presence of an additional ORF 184 bp downstream of hupL, showing a high degree of sequence identity with a gene encoding an uptake- hydrogenase-specific enclopepticlase (hupW) in other cyanobacteria. In addition, the transcription start point was identified 238 bp upstream of the hupS translational start. RT-PCR experiments revealed that hupW is co-transcribed with the uptake hydrogenase structural genes in Gloeothece sp. ATCC 27152. In addition, Northern hybridizations clearly showed that hupSL Ware transcribed under nitrogen fixing conditions, but not in the presence of combined nitrogen. A putative NtcA binding site was identified in the promoter region upstream of hupS, centred at -41(.)5 bp with respect to the transcription start point. Electrophoretic retardation of a labelled DNA fragment (harbouring the putative NtcA-binding motif) was significantly affected by an Escherichia coli cell-free extract containing overexpressed NtcA, suggesting that NtcA is involved in the transcriptional regulation of hupSLW.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 9