Molecular cloning, phylogeny and localization of AgNHA1: the first Na+/H+ antiporter (NHA) from a metazoan, Anopheles gambiae.
Mark R. Rheault,Bernard A. Okech,Stephen B. W. Keen,Melissa M. Miller,Ella A. Meleshkevitch,Paul J. Linser,Dmitri Y. Boudko,William R. Harvey +7 more
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The tissue localization of AgNHA1 suggests that it plays a key role in maintaining the characteristic longitudinal pH gradient in the lumen of the alimentary canal of An.Abstract:
SUMMARY We have cloned a cDNA encoding a new ion transporter from the alimentary
canal of larval African malaria mosquito, Anopheles gambiae Giles
sensu stricto . Phylogenetic analysis revealed that the corresponding
gene is in a group that has been designated NHA, and which includes
(Na + or K + )/H + antiporters; so the novel
transporter is called AgNHA1. The annotation of current insect genomes shows
that both AgNHA1 and a close relative, AgNHA2, belong to the cation proton
antiporter 2 (CPA2) subfamily and cluster in an exclusive clade of genes with
high identity from Aedes aegypti , Drosophila melanogaster ,
D. pseudoobscura , Apis mellifera and Tribolium
castaneum . Although NHA genes have been identified in all phyla for which
genomes are available, no NHA other than AgNHA1 has previously been cloned,
nor have the encoded proteins been localized or characterized. The AgNHA1 transcript was localized in An. gambiae larvae by
quantitative real-time PCR (qPCR) and in situ hybridization. AgNHA1
message was detected in gastric caeca and rectum, with much weaker
transcription in other parts of the alimentary canal. Immunolabeling of whole
mounts and longitudinal sections of isolated alimentary canal showed that
AgNHA1 is expressed in the cardia, gastric caeca, anterior midgut, posterior
midgut, proximal Malpighian tubules and rectum, as well as in the
subesophageal and abdominal ganglia. A phylogenetic analysis of NHAs and KHAs indicates that they are
ubiquitous. A comparative molecular analysis of these antiporters suggests
that they catalyze electrophoretic alkali metal ion/hydrogen ion exchanges
that are driven by the voltage from electrogenic H + V-ATPases. The
tissue localization of AgNHA1 suggests that it plays a key role in maintaining
the characteristic longitudinal pH gradient in the lumen of the alimentary
canal of An. gambiae larvae.read more
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