Title: A novel aquaporin 3 in killifish (Fundulus heteroclitus) is not an arsenic channel.
Authors: Jung, Dawoon; MacIver, Bryce; Jackson, Brian P; Barnaby, Roxanna; Sato, J Denry; Zeidel, Mark L; Shaw, Joseph R; Stanton, Bruce A
Published In Toxicol Sci, (2012 May)
Abstract: The Atlantic killifish (Fundulus heteroclitus) is a model environmental organism that has an extremely low assimilation rate of environmental arsenic. As a first step in elucidating the mechanism behind this phenomenon, we used quantitative real-time PCR to identify aquaglyceroporins (AQPs), which are arsenite transporters, in the killifish gill. A novel homolog killifish AQP3 (kfAQP3a) was cloned from the killifish gill, and a second homolog was identified as the consensus from a transcriptome database (kfAQP3b). The two were 99% homologous to each other, 98% homologous to a previously identified killifish AQP3 from embryos (kfAQP3ts), and 78% homologous to hAQP3. Expression of kfAQP3a in Xenopus oocytes significantly enhanced water, glycerol, and urea transport. However, kfAQP3a expressed in HEK293T cells did not transport significant amounts of arsenic. All sequence motifs thought to confer the ability of AQP3 to transport solutes were conserved in kfAQP3a, kfAQP3b, and kfAQP3ts; however, the C-terminal amino acids were different in kfAQP3a versus the other two homologs. Replacement of the three C-terminal amino acids of kfAQP3 (GKS) with the three C-terminal amino acids of kfAQP3b and kfAQP3ts (ANC) was sufficient to enable kfAQP3a to robustly transport arsenic. Thus, the C-terminus of kfAQP3b and kfAQP3ts confers arsenic selectivity in kfAQP3. Moreover, kfAQP3a, the only AQP expressed in killifish gill, is the first aquaglyceroporin identified that does not transport arsenic, which may explain, in part, why killifish poorly assimilate arsenic and are highly tolerant to environmental arsenic.
PubMed ID: 22323512
MeSH Terms: Animals; Aquaporin 3/chemistry; Aquaporin 3/genetics; Aquaporin 3/metabolism*; Arsenites/metabolism; Arsenites/toxicity*; Biological Transport/drug effects; Biological Transport/physiology; Cloning, Molecular; Disease Models, Animal; Fish Proteins/genetics; Fish Proteins/metabolism*; Fundulidae/physiology*; Gene Expression Regulation/drug effects; Gills/chemistry; HEK293 Cells/drug effects; HEK293 Cells/metabolism; Humans; Oocytes/metabolism; RNA, Messenger/metabolism; Real-Time Polymerase Chain Reaction; Sodium Compounds/metabolism; Sodium Compounds/toxicity*; Species Specificity; Structure-Activity Relationship; Water Pollutants, Chemical/metabolism; Water Pollutants, Chemical/toxicity*; Xenopus/physiology