Role of N-terminal hydrophobic region in modulating the subcellular localization and enzyme activity of the bisphosphate nucleotidase from Debaryomyces hansenii.
3', 5'-Bisphosphate nucleotidase is a ubiquitous enzyme that converts 3'-phosphoadenosine-5'-phosphate to adenosine-5'-phosphate and inorganic phosphate. These enzymes are highly sensitive to sodium and lithium and, thus, perform a crucial rate-limiting metabolic step during salt stress in yeast. Recently, we have identified a bisphosphate nucleotidase gene (DHAL2) from the halotolerant yeast Debaryomyces hansenii. One of the unique features of Dhal2p is that it contains an N-terminal 54-amino-acid-residue hydrophobic extension. In this study, we have shown that Dhal2p exists as a cytosolic as well as a membrane-bound form and that salt stress markedly influences the accumulation of the latter form in the cell. We have demonstrated that the N-terminal hydrophobic region was necessary for the synthesis of the membrane-bound isoform. It appeared that an alternative translation initiation was the major mechanism for the synthesis of these two forms. Moreover, the two forms exhibit significant differences in their substrate specificity. Unlike the cytosolic form, the membrane-bound form showed very high activity against inositol-1,4-bisphosphate. Thus, the present study for the first time reports the existence of multiple forms of a bisphosphate nucleotidase in any organism.
Institute of Microbial Technology, Sector 39A, Chandigarh 160 036, India.
Monika
M
Aggarwal
Alok K
AK
Mondal
D000595:Amino Acid Sequence; D018387:Codon, Initiator / Q000235:genetics; D020869:Gene Expression Profiling; D005800:Genes, Fungal / Q000235:genetics; D005822:Genetic Vectors; D057927:Hydrophobic and Hydrophilic Interactions; D008566:Membranes / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009708:Nucleotidases / Q000737:chemistry* / Q000378:metabolism*; D010442:Peptide Chain Initiation, Translational / Q000235:genetics; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D021381:Protein Transport; D004718:Saccharomycetales / Q000166:cytology* / Q000201:enzymology* / Q000254:growth & development; D012492:Salts / Q000378:metabolism
D018387:Codon, Initiator; D012492:Salts; D009708:Nucleotidases; D010744:Phosphoric Monoester Hydrolases; C053087:inositol-1,4-bisphosphate 1-phosphatase; C053354:bisphosphoadenylate 3'-nucleotidase