title: An SLC6 transporter of the novel B(0,)- system aids in absorption and detection of nutrient amino acids in Caenorhabditis elegans.
abstract: Nutrient amino acid transporters (NATs) of solute carrier family 6 (SLC6) mediate uptake of essential amino acids in mammals and insects. Phylogenomic analysis of the Caenorhabditis elegans (Ce) SLC6 family identifies five genes paralogous to an insect-specific NAT subfamily. Here we cloned and characterized the first representative of the identified nematode-specific transporters, SNF-5. SNF-5 mediates broad spectrum cation-coupled transport of neutral amino acids with submillimolar affinities and stoichiometry of 1 AA:1 Na(+), except for 1 l-Pro:2 Na(+). Unexpectedly, it transports acidic l-Glu(-) and l-Asp(-) (1 AA(-):3 Na(+)), revealing it to be the first member of a new B(0,-) system among characterized SLC6 transporters. This activity correlates with a unique positively charged His(+) 377 in the substrate-binding pocket. snf-5 promoter-driven enhanced green fluorescent protein labels intestinal cells INT1-9 and three pairs of amphid sensory neurons: ASI, ADF and ASK. These cells are intimately involved in control of dauer diapause, development, metabolism and longevity. The snf-5 deletion mutants do not show apparent morphological disorders, but increase dauer formation while reducing dauer maintenance upon starvation. Overall, the present study characterized the first nematode-specific NAT and revealed important structural and functional aspects of this transporter. In addition to the predictable role in alimentary amino acid absorption, our results indicate possible neuronal roles of SNF-5 as an amino acid provider to specific neuronal functions, including sensing of amino acid availability.
authors.affiliation: The Department of Physiology and Biophysics of the Rosalind Franklin University of Medicine and Science, Chicago Medical School, North Chicago, IL 60064, USA.
authors.forename: Ryan
authors.initials: R
authors.lastname: Metzler
authors.forename: Ella A
authors.initials: EA
authors.lastname: Meleshkevitch
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Fox
authors.forename: Hongkyun
authors.initials: H
authors.lastname: Kim
authors.forename: Dmitri Y
authors.initials: DY
authors.lastname: Boudko
mesh_terms: D000042:Absorption; D000595:Amino Acid Sequence; D026921:Amino Acid Transport Systems, Neutral / Q000737:chemistry / Q000378:metabolism*; D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D017173:Caenorhabditis elegans / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry / Q000378:metabolism*; D003001:Cloning, Molecular; D055724:Electrophysiological Phenomena; D017353:Gene Deletion; D016678:Genome / Q000235:genetics; D007477:Ions; D007553:Isotope Labeling; D007700:Kinetics; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010641:Phenotype; D010802:Phylogeny; D016415:Sequence Alignment; D027981:Symporters / Q000737:chemistry / Q000378:metabolism*
chemical_list: D026921:Amino Acid Transport Systems, Neutral; D000596:Amino Acids; D029742:Caenorhabditis elegans Proteins; D007477:Ions; C586048:SNF-5 protein, C elegans; D027981:Symporters
keywords: ADF; ASI; ASK; B0,- system; SLC6; amphid sensory neurons; dauer; embryonic arrest; glutamate; snf-5; transceptor