Cloning and molecular characterization of GroESL heat-shock operon in methylotrophic bacterium Methylovorus Sp. strain SS1 DSM 11726.
The groESL bicistronic operon of a restricted facultative methylotrophic bacterium Methylovorus sp. strain SS1 DSM 11726 was cloned and characterized. It was found to consist of two ORFs encoding proteins with molecular masses of 11,395 and 57,396 daltons, which showed a high degree of homology to other bacterial GroES and GroEL proteins. The genes were clustered in the transcription order groES-groEL. Northern blot analyses suggested that the groESL operon is transcribed as a bicistronic 2.2-kb mRNA, the steady-state level of which was markedly increased by temperature elevation. Primer extension analysis demonstrated one potential transcription start site preceding the groESL operon, which is located 100 bp upstream of the groES start codon. The transcription start site was preceded by a putative promoter region highly homologous to the consensus sequences of Escherichia coli sigma 32-type heat shock promoter, which functioned under both normal and heat shock conditions in E. coli. Heat shock mRNA was maximally produced by Methylovorus sp. strain SS1 approximately 10 min after increasing the temperature from 30 to 42 degrees C. The groESL operon was also induced by hydrogen peroxide or salt shock.
Eom
Chi Yong
CY
Department of Biology, Yonsei University, Seoul 120-749, Korea.
Kim
Eungbin
E
Ro
Young Tae
YT
Kim
Si Wouk
SW
Kim
Young Min
YM
D001419:Bacteria / Q000378:metabolism*; D018835:Chaperonin 10 / Q000737:chemistry*; D018834:Chaperonin 60 / Q000737:chemistry*; D003062:Codon; D017931:DNA Primers / Q000737:chemistry; D015964:Gene Expression Regulation, Bacterial; D018869:Heat-Shock Response / Q000235:genetics; D006861:Hydrogen Peroxide / Q000737:chemistry; D008954:Models, Biological; D009876:Operon; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012492:Salts / Q000737:chemistry; D013696:Temperature; D014158:Transcription, Genetic
D018835:Chaperonin 10; D018834:Chaperonin 60; D003062:Codon; D017931:DNA Primers; D012492:Salts; D006861:Hydrogen Peroxide