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Rezumat articol ediţie STUDIA UNIVERSITATIS BABEŞ-BOLYAI
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Rezumat: Proteins of Lsm family are key players in RNA metabolism. However, archaeal homologs of these proteins (Like-Sm proteins) have been less studied than in the rest of the domains of life. H. mediterranei encodes a single lsm gene, which overlaps by four nucleotides with the rl37e gene. The gene environment anaylsis has revealed that it is highly conserved in Archaea. In most of the archaeal genomes analysed, the rpl37e gene follows the lsm gene. In 56 genomes, the two genes overlap and in 21 genomes they are widely spaced, so the cotranscription could be assumed. Expression and co-transcription with the rl37e gene were analysed by RT-PCR in complex medium, defined medium with nitrate or ammonium as nitrogen source, and nitrogen starvation at different times. The lsm and rl37e genes are expressed in all conditions, but only cotranscribe under nitrogen starvation conditions. Gene expression microarray shows that the lsm gene presents non-significant expression differences in different nitrogen sources. To determine the Lsm function, deletion mutants of lsm1 gene (HM26-Δlsm) and Sm1 domain (HM26-ΔSm1) were generated. These mutants were characterised in the presence of various sources of nitrogen and complex medium. Moreover, swarming assays were carried out and the cell morphology was analysed in the different growth phases in the same culture media. Finally, heat shock, oxidative stress and ethanol stress assays were performed. The deletion of the lsm gene and Sm1 domain are viable, resulting in a pleiotropic phenotype, indicating that the haloarchaeal Lsm protein is involved in many cellular processes.
Keywords: deletion mutants, haloarchaea, Lsm protein, microarray, RT-PCR.
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