Molecular cloning and characterization of a complementary DNA encoding sperm tail protein SHIPPO 1.
Formation of the tail in developing sperm is a complex process involving the organization of the axoneme, transport of periaxonemal proteins from the cytoplasm to the tail, and assembly of the outer dense fibers and fibrous sheath. Although detailed morphological descriptions of these events are available, the molecular mechanisms remain to be fully elucidated. We have isolated a new gene, named shippo 1, from a haploid germ cell-specific cDNA library of mouse testis, and also its human orthologue (h-shippo 1). The isolated cDNA is 1.2 kilobases long, carrying a 762-base pair open reading frame that encodes SHIPPO 1, a sperm protein predicted to consist of 254 amino acids. The amino acid sequence includes 6 Pro-Gly-Pro repeats, which are also present in the human orthologue protein (hSHIPPO 1) as well as in 2 other newly reported proteins of Drosophila melanogaster. Transcription of shippo 1 is exclusively observed in haploid germ cells. Antibody raised against SHIPPO 1 identified a testis-specific M(r) 32 x 10(-3) band in Western blot analysis. The protein was further localized in the flagella of the elongated spermatids and along the entire length of the tail in mature sperm. SHIPPO 1 in sperm is resistant to treatment with nonionic detergents and coextracted with the cytoskeletal core proteins of the mouse sperm tail.
Egydio de Carvalho
Carlos
C
Department of Science for Laboratory Animal Experimentation, Research Institute for Microbial Diseases, Osaka University, Suita City, Osaka 565-0871, Japan.
Tanaka
Hiromitsu
H
Iguchi
Naoko
N
Ventelä
Sami
S
Nojima
Hiroshi
H
Nishimune
Yoshitake
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002458:Cell Fractionation; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics*; D015870:Gene Expression; D015723:Gene Library; D006238:Haploidy; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009928:Organ Specificity; D029589:Seminal Plasma Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D017422:Sequence Analysis, DNA; D013082:Sperm Tail / Q000737:chemistry*; D013094:Spermatozoa / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D013737:Testis / Q000737:chemistry
D018076:DNA, Complementary; C455114:Odf3 protein, mouse; D029589:Seminal Plasma Proteins