Identification of genes regulated by changing salinity in the deep-sea bacterium Shewanella sp. WP3 using RNA arbitrarily primed PCR.

The differential gene transcription of a deep-sea bacterium Shewanella sp. WP3 in response to changing salinity was analyzed by RNA fingerprinting using arbitrarily primed PCR (RAP-PCR). Ninety primer sets were used to scan two different RNA pools derived from cultures of 1% and 7% NaCl concentrations. Forty-three putative differential-expressed fragments were identified, cloned, and sequenced. Six out of the 43 fragments were confirmed to be truly differentially transcribed in terms of changing salinity. The deduced amino acid sequences of the six gene fragments showed highest identities (66-96%) with ribosomal protein L24, ATP binding protein, and chaperon protein HscA of Shewanella oneidensis MR-1 (Y6, Y9, and Y29); isocitrate lyase of Pseudomonas aeruginosa (Y15); peptidylprolyl cis-trans isomerase of Shewanella sp. SIB1 (Y21), glutamine synthetase of Shewanella violacea (Y25), respectively. Four genes (Y6, Y15, Y21, and Y25) were up regulated in 7% NaCl, while the other two (Y9 and Y29) contained more abundant transcripts in 1% NaCl. The data suggested that strategies involved in controlling protein synthesis, protein folding and/or trafficking, glutamate concentration, fatty acid metabolism, and substance transporting were used for salt adaptation in Shewanella sp. WP3. The expression patterns of the six genes in response to transient stress shocks including salt shock (3% NaCl shift to 12%), cold shock (15 degrees C shift to 0 degrees C), and high-hydrostatic pressure shock (0.1 MPa shift to 50 MPa) were further examined. Y29 encoding the putative HscA chaperon protein was indicated to be involved in adaptation of all the stresses tested.