Cloning and sequencing of SOB3, a human gene coding for a sperm protein homologous to an antimicrobial protein and potentially involved in zona pellucida binding.
We have previously characterized an 18-19 kDa cationic protein, SOB3, that was detected in the epididymis and localized within the acrosome and on the neck region of human spermatozoa. We suggested that it is involved in secondary sperm binding to the zona pellucida. The present study describes its purification to homogeneity by preparative electrophoresis and non-equilibrium pH gradient electrophoresis. Degenerate primers deduced from microsequencing were used to amplify a specific fragment from human epididymal RNA by reverse transcription-polymerase chain reaction (RT-PCR). This 164 bp fragment was extended by 5' and 3'-RACE to obtain the 548 bp full length cDNA. The open reading frame encodes a 170 amino acid protein. SOB3 is a single copy gene. It is 98% identical to prepro-FALL39 and 100% identical to CAP18, two human genes which were initially identified by screening a human bone marrow (lambda)gt11 library, and which encode an antimicrobial protein. Northern blots of human tissues revealed a 1 kb transcript in corpus and cauda epididymis only, while RT-PCR showed presence of the mRNA in the three epididymal regions and also in round spermatids. The above results suggest that SOB3 has two roles in sperm protection and fertilization, depending on its dual origin and final sperm localization.
Hammami-Hamza
S
S
INSERM U 355, Maturation Gamétique et Fécondation, Institut Paris-sud sur les Cytokines, 32 rue des Carnets, 92140 Clamart, France.
Doussau
M
M
Bernard
J
J
Rogier
E
E
Duquenne
C
C
Richard
Y
Y
Lefèvre
A
A
Finaz
C
C
D000595:Amino Acid Sequence; D000818:Animals; D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides / Q000235:genetics*; D001483:Base Sequence; D054804:Cathelicidins; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005260:Female; D006801:Humans; D007633:Keratins / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D012313:RNA; D017422:Sequence Analysis, DNA; D013084:Sperm-Ovum Interactions / Q000502:physiology; D013094:Spermatozoa / Q000378:metabolism / Q000502:physiology*; D014018:Tissue Distribution; D015044:Zona Pellucida / Q000502:physiology*
D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D054804:Cathelicidins; D018076:DNA, Complementary; D012313:RNA; D007633:Keratins