Heme uptake genes in human and fish isolates of Photobacterium damselae: existence of hutA pseudogenes

Abstract

The marine bacterium Photobacterium damselae includes strains classified into two distinct subspecies, namely subsp. damselae and subsp. piscicida, which have been reported to cause disease in a variety of marine animals and in humans. P. damselae strains utilize heme compounds as sole iron sources. In the present study, ten potential heme uptake and utilization genes are described in P. damselae subsp. damselae and subsp. piscicida. One gene cluster includes the genes coding for putative proteins HutZ, HutX and HutW; TonB, ExbB and ExbD, the three components of the TonB system; HutB, the putative periplasmic binding protein; HutC, the putative inner membrane permease; and HutD, the putative ABC-transporter ATP-ase. A gene coding for HutA, the outer membrane heme receptor, has also been identified, but it is not linked to the rest of the heme transport genes. RT-PCR analyses showed that heme uptake genes are arranged in three iron-regulated transcriptional units. A plasmid carrying the gene for the heme receptor HutA in combination with a plasmid carrying tonBexbBDhutBCD genes conferred to Escherichia coli 101ESD (ent) the ability to use heme and hemoglobin as iron sources. The hutA gene was present in strains isolated from humans and a variety of fish species, but it was shown to be interrupted in some subsp. piscicida strains, constituting a pseudogene. This is the first description of a heme-uptake system in a Photobacterium species, and shows some structural and functional similarities to heme-uptake systems reported in other gram-negative bacteria.