Isolation, sequence and characterization of the maltose-regulated mlrA gene from the hyperthermophilic archaeum...

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.