Anaerobic growth, a property horizontally transferred by an Hfr-like mechanism among extreme thermophiles.
Despite the fact that the extreme thermophilic bacteria belonging to the genus Thermus are classified as strict aerobes, we have shown that Thermus thermophilus HB8 (ATCC 27634) can grow anaerobically when nitrate is present in the growth medium. This strain-specific property is encoded by a respiratory nitrate reductase gene cluster (nar) whose expression is induced by anoxia and nitrate (S. Ramírez-Arcos, L. A. Fernández-Herrero, and J. Berenguer, Biochim. Biophys. Acta, 1396:215-1997). We show here that this nar operon can be transferred by conjugation to an aerobic Thermus strain, enabling it to grow under anaerobic conditions. We show that this transfer takes place through a DNase-insensitive mechanism which, as for the Hfr (high frequency of recombination) derivatives of Escherichia coli, can also mobilize other chromosomal markers in a time-dependent way. Three lines of evidence are presented to support a genetic linkage between nar and a conjugative plasmid integrated into the chromosome. First, the nar operon is absent from a plasmid-free derivative and from a closely related strain. Second, we have identified an origin for autonomous replication (oriV) overlapping the last gene of the nar cluster. Finally, the mating time required for the transfer of the nar operon is in good agreement with the time expected if the transfer origin (oriT) were located nearby and downstream of nar.
Ramírez-Arcos
S
S
Centro de Biología Molecular Severo Ochoa, UAM-CSIC, Universidad Autónoma de Madrid, Spain.
Fernández-Herrero
L A
LA
Marín
I
I
Berenguer
J
J
D000595:Amino Acid Sequence; D000693:Anaerobiosis; D001483:Base Sequence; D003227:Conjugation, Genetic; D003470:Culture Media; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D005810:Multigene Family; D050901:Nitrate Reductase; D009565:Nitrate Reductases / Q000235:genetics; D009566:Nitrates / Q000378:metabolism; D009876:Operon; D010957:Plasmids / Q000235:genetics; D018741:Replication Origin; D015183:Restriction Mapping; D016963:Thermus thermophilus / Q000235:genetics* / Q000254:growth & development* / Q000378:metabolism
D003470:Culture Media; D004269:DNA, Bacterial; D009566:Nitrates; D009565:Nitrate Reductases; D050901:Nitrate Reductase