Isolation, sequence and characterization of the maltose-regulated mlrA gene from the hyperthermophilic archaeum Pyrococcus furiosus.
The mlrA (maltose regulated) gene from the hyperthermophilic archaeum Pyrococcus furiosus was identified from a family of clones whose expression was influenced by the presence of maltose in the medium. Sequencing of the 2276 bp of DNA containing mlrA and flanking regions revealed a 753-amino-acid (aa) (88 kDa) open reading frame (ORF). The ORF is preceded by a bacterial-like ribosome-binding site. The deduced product shared extensive homology with pyruvate dikinases (PDK) from both eukaryal and eubacterial sources (35-61% similarity) and the signature domains characteristic of this class of proteins were present. Northern blot experiments demonstrated the presence of an approx. 2.4-kb transcript in P. furiosus extracts, corresponding in length to that expected from expression of mlrA. P. furiosus cultures grown in the presence of maltose were found to contain approx. 5-10-fold greater mlrA mRNA than those grown without maltose. Initiation of transcription under both cultural conditions occurred at the same transcription start point (tsp), 23 bp downstream from a putative BoxA promoter element.
Robinson
K A
KA
Center of Marine Biotechnology, University of Maryland Biotechnology Institute, Baltimore 21202.
Schreier
H J
HJ
D000595:Amino Acid Sequence; D001105:Archaea / Q000235:genetics* / Q000378:metabolism; D019843:Archaeal Proteins; D001419:Bacteria / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D005798:Genes, Bacterial; D006358:Hot Temperature; D008320:Maltose / Q000378:metabolism; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic
D019843:Archaeal Proteins; D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; C091619:MlrA protein, Pyrococcus furiosus; D011994:Recombinant Proteins; D008320:Maltose