Characterization of the glutamate dehydrogenase gene and its regulation in a euryhaline copepod.
Glutamate dehydrogenase (GDH) plays a key role in the metabolism of free amino acids (FAA) in crustaceans and other metazoans. Glutamate synthesized by GDH via reductive amination is the amino group donor for alanine synthesis and the precursor required for proline synthesis. Since both proline and alanine are important intracellular osmolytes in many marine invertebrates, GDH has been widely implicated as playing a central role in response to hyperosmotic stress in these organisms. We have isolated the gene encoding a GDH homolog from the euryhaline copepod Tigriopus californicus and examined the regulation of GDH under salinity stress. The gene encodes a protein of 557 residues with 76% amino acid identity with Drosophila melanogaster GDH. The gene encodes an N-terminal mitochondrial signal sequence peptide. Only a single intron of 71 bp was found in the GDH gene in T. californicus when genomic sequences and cDNA sequences were compared. The levels of GDH mRNA do not increase during hyperosmotic stress in this copepod. The effects of salt and hyperosmotic stress on GDH enzyme activity were also investigated. GDH activities decrease with increasing NaCl concentrations in in vitro enzyme assays, while live animals exposed to hyperosmotic stress showed no change in GDH enzyme activities. Combined, these results indicate that GDH transcription and enzyme activity do not appear to function in the regulation of alanine and proline accumulation during hyperosmotic stress in T. californicus. The manner in which this important physiological process is regulated remains unknown.
Willett
Christopher S
CS
Department of Biology, University of North Carolina, CB#3280 Coker Hall, Chapel Hill, NC 27599-3280, USA. willett4@email.unc.edu
Burton
Ronald S
RS
D000595:Amino Acid Sequence; D000818:Animals; D033342:Copepoda / Q000201:enzymology* / Q000235:genetics; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D005969:Glutamate Dehydrogenase / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009994:Osmolar Concentration; D010802:Phylogeny; D012333:RNA, Messenger / Q000378:metabolism; D016415:Sequence Alignment
D029721:Drosophila Proteins; D012333:RNA, Messenger; D018698:Glutamic Acid; D005969:Glutamate Dehydrogenase