Characterization and transcriptional analysis of hupSLW in Gloeothece sp. ATCC 27152: an uptake hydrogenase from a unicellular cyanobacterium
| Autor: | Paula Tamagnini, Paulo J. Oliveira, Elsa Leitão, Pedro Moradas-Ferreira, Fredrik Oxelfelt |
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| Rok vydání: | 2004 |
| Předmět: |
DNA
Bacterial Hydrogenase Nitrogen Molecular Sequence Data Electrophoretic Mobility Shift Assay Biology Cyanobacteria medicine.disease_cause Microbiology Bacterial Proteins Nitrogen Fixation Gene Order medicine Amino Acid Sequence RNA Messenger Promoter Regions Genetic Gene Escherichia coli Base Sequence Sequence Homology Amino Acid Reverse Transcriptase Polymerase Chain Reaction Structural gene Nucleic acid sequence Biological Transport Promoter Sequence Analysis DNA Blotting Northern Genetic translation DNA-Binding Proteins RNA Bacterial Open reading frame Biochemistry Genes Bacterial Transcription Initiation Site Oxidoreductases Sequence Alignment |
| Zdroj: | Microbiology. 150:3647-3655 |
| ISSN: | 1465-2080 1350-0872 |
| DOI: | 10.1099/mic.0.27248-0 |
| Popis: | 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 endopeptidase (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 hupSLW are 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. |
| Databáze: | OpenAIRE |
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