VHY, a novel myristoylated testis-restricted dual specificity protein phosphatase related to VHX.
The human DUSP15 gene encodes an uncharacterized 235-amino acid member of the subfamily of small dual specificity protein phosphatases related to the Vaccinia virus VH1 phosphatase. Similar to VHR-related MKPX (VHX) (DUSP22), the predicted protein has an N-terminal myristoylation recognition sequence, and we show here that both are indeed modified by the attachment of a myristate to Gly-2. In recognition of this relatedness to VHX, we refer to the DUSP15-encoded protein as VH1-related member Y (VHY). We report that VHY is expressed at high levels in the testis and barely detectable levels in the brain, spinal cord, and thyroid. A VHY-specific antiserum detected a protein with an apparent molecular mass of 26 kDa, and histochemical analysis showed that VHY was readily detectable in pachytene spermatocytes (midstage of meiotic division I) and round spermatids and weakly in Leydig cells (somatic cells outside of the seminiferous tubules). When expressed in 293T or NIH-3T3 cells, VHY was concentrated at the plasma membrane with some staining of vesicular structures in the Golgi region. Mutation of the myristoylation site Gly-2 abrogated membrane location. Finally, we demonstrate that VHY is an active phosphatase in vitro. We conclude that VHY is a new member of a subgroup of myristoylated VH1-like small dual specificity phosphatases.
Alonso
Andres
A
Program of Signal Transduction, Cancer Research Center, The Burnham Institute, La Jolla, California 92037, USA.
Narisawa
Sonoko
S
Bogetz
Jori
J
Tautz
Lutz
L
Hadzic
Radinka
R
Huynh
Huong
H
Williams
Scott
S
Gjörloff-Wingren
Anette
A
Bremer
Meire C D
MC
Holsinger
Leslie J
LJ
Millan
José L
JL
Mustelin
Tomas
T
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D015152:Blotting, Northern; D015139:Blotting, Southern; D002460:Cell Line; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D054637:Dual-Specificity Phosphatases; D004622:Embryo, Mammalian / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D005998:Glycine / Q000737:chemistry; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D048031:JNK Mitogen-Activated Protein Kinases; D048670:MAP Kinase Kinase 4; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism; D054639:Mitogen-Activated Protein Kinase Phosphatases; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009227:Myristic Acids / Q000737:chemistry; D041681:NIH 3T3 Cells; D009596:Nitrophenols / Q000737:chemistry; D009943:Organophosphorus Compounds / Q000737:chemistry; D010749:Phosphoprotein Phosphatases / Q000737:chemistry*; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D017434:Protein Structure, Tertiary; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012097:Repressor Proteins / Q000737:chemistry* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013087:Spermatids / Q000378:metabolism; D013737:Testis / Q000378:metabolism; D014162:Transfection; D014616:Vaccinia virus / Q000378:metabolism
D018076:DNA, Complementary; D009227:Myristic Acids; D009596:Nitrophenols; D009943:Organophosphorus Compounds; D012333:RNA, Messenger; D012097:Repressor Proteins; C008644:nitrophenylphosphate; D005982:Glutathione Transferase; D048031:JNK Mitogen-Activated Protein Kinases; D048670:MAP Kinase Kinase 4; D020929:Mitogen-Activated Protein Kinase Kinases; D054639:Mitogen-Activated Protein Kinase Phosphatases; D010749:Phosphoprotein Phosphatases; D010744:Phosphoric Monoester Hydrolases; C442824:DUSP22 protein, human; D054637:Dual-Specificity Phosphatases; C522152:Dusp22 protein, mouse; D017027:Protein Tyrosine Phosphatases; D005998:Glycine