Cloning, sequencing, and characterization of ribosomal protein and RNA polymerase genes from the region analogous to the alpha-operon of escherichia coli in halophilic archaea, halobacterium halobium.
A determination was made of the nucleotide sequence of the 3215-bp region of a ribosomal protein gene cluster (HS13, HS4, HS11, and HeL18), RNA polymerase (RNA poly D), and tRNA genes (tRNAser and tRNAarg) of halophilic Archaea Halobacterium halobium, which is analogous to the alpha-operon of Escherichia coli (tRNAser-HS13-HS4-HS11-RNA poly D-tRNAarg-HeL18). The seven-gene string was preceded by a pseudoknot-like structure similar to the proposed S4 ribosomal protein binding site of the alpha-operon mRNA leader in E. coli. Using an inducible expression system H. halobium HS4 was produced in large amounts in E. coli, and immunoblot analysis showed the S4 to constitute a 21-kDa polypeptide component of the ribosome. Analysis of the deduced amino acids sequence revealed that the HS13, HS4, and HS11 sequences including the RNA polymerase subunit are more similar to their eukaryotic than to their bacterial counterparts. HeL18, located downstream of the gene cluster analogous to the E. coli alpha-operon (S13-S11-S4-RNA poly D-L17), was similar to both the eukaryotic (eL18) and eubacterial ribosomal protein L15 located in the spc-operon, but not to L17 positioned as the terminal gene of the bacterial alpha-operon.
Sano
K
K
School of Bioresources, Hiroshima Prefectural University, Shobara City, Hiroshima, 727-0023, Japan.
Taguchi
A
A
Furumoto
H
H
Uda
T
T
Itoh
T
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D019641:DNA, Archaeal / Q000032:analysis / Q000737:chemistry; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D019766:Genes, Archaeal; D019617:Halobacterium salinarum / Q000235:genetics*; D008969:Molecular Sequence Data; D005810:Multigene Family; D009690:Nucleic Acid Conformation; D009876:Operon; D010802:Phylogeny; D012347:RNA, Transfer, Arg / Q000235:genetics; D012362:RNA, Transfer, Ser / Q000235:genetics; D012269:Ribosomal Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D017386:Sequence Homology, Amino Acid
D019641:DNA, Archaeal; D012347:RNA, Transfer, Arg; D012362:RNA, Transfer, Ser; D012269:Ribosomal Proteins; C032493:ribosomal protein L17; C029609:ribosomal protein L18; C031798:ribosomal protein S4; D012321:DNA-Directed RNA Polymerases