Sequence, expression and evolution of the globins of the parasitic nematode Nippostrongylus brasiliensis

The globins of the nematode parasite Nippostrongylus brasiliensis have oxygen affinities 100-fold higher than the rodent host's haemoglobins. Two isoforms are found, one located in the cuticle, and the other in the body of the nematode. Both isoforms have been cloned and analysed for clues as to function and evolution. The body globin isoform is first expressed upon invasion of the mammalian host. The abundant cuticular globin is expressed only by adult nematodes in the gut, and differs significantly from the body globin. Both globins are found as trans-spliced mRNAs: the developmental pattern of expression of the mRNA parallels the protein expression. The pattern of the nematode globin genes is complex. Comparison with other nematode globin sequences suggests that N. brasiliensis is more closely related to Caenorhabditis elegans than to ascarid species. At least two gene duplication events are predicted: gene duplication preceded the radiation of the important vertebrate-parasitic strongylid nematode species. Both N. brasiliensis globins have a central intron the exact position of which suggests that it arose from an independent insertion event in the strongylid-rhabditid line. The globins have been expressed in Escherichia coli as functional holenzymes as a prelude to studies to elucidate the origin of their extraordinary oxygen affinity.