Characterization of gametogenetin 1 (GGN1) and its potential role in male fertility through the interaction with the ion channel regulator, cysteine-rich secretory protein 2 (CRISP2) in the sperm tail.
Cysteine-rich secretory protein 2 (CRISP2) is a testis-enriched protein localized to the sperm acrosome and tail. CRISP2 has been proposed to play a critical role in spermatogenesis and male fertility, although the precise function(s) of CRISP2 remains to be determined. Recent data have shown that the CRISP domain of the mouse CRISP2 has the ability to regulate Ca(2+) flow through ryanodine receptors (RyR) and to bind to MAP kinase kinase kinase 11 (MAP3K11). To further define the biochemical pathways within which CRISP2 is involved, we screened an adult mouse testis cDNA library using a yeast two-hybrid assay to identify CRISP2 interacting partners. One of the most frequently identified CRISP2-binding proteins was gametogenetin 1 (GGN1). Interactions occur between the ion channel regulatory region within the CRISP2 CRISP domain and the carboxyl-most 158 amino acids of GGN1. CRISP2 does not bind to the GGN2 or GGN3 isoforms. Furthermore, we showed that Ggn1 is a testis-enriched mRNA and the protein first appeared in late pachytene spermatocytes and was up-regulated in round spermatids before being incorporated into the principal piece of the sperm tail where it co-localized with CRISP2. These data along with data on RyR and MAP3K11 binding define the CRISP2 CRISP domain as a protein interaction motif and suggest a role for the GGN1-CRISP2 complex in sperm tail development and/or motility.
Jamsai
Duangporn
D
The Centre for Reproduction and Development and The Centre for Urological Research, Monash Institute of Medical Research, Monash University, 27-31 Wright Street, Clayton, Melbourne, Victoria 3168, Australia. duangporn.jamsai@med.monash.edu.au
Bianco
Deborah M
DM
Smith
Stephanie J
SJ
Merriner
Donna J
DJ
Ly-Huynh
Jennifer D
JD
Herlihy
Amy
A
Niranjan
Birunthi
B
Gibbs
Gerard M
GM
O'Bryan
Moira K
MK
D000177:Acrosome / Q000737:chemistry / Q000378:metabolism; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern / Q000379:methods; D015153:Blotting, Western / Q000379:methods; D015815:Cell Adhesion Molecules; D003001:Cloning, Molecular; D005818:Genetic Engineering; D006023:Glycoproteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry; D008297:Male; D008565:Membrane Proteins; D051379:Mice; D008969:Molecular Sequence Data; D011485:Protein Binding; D013081:Sperm Motility / Q000502:physiology; D013082:Sperm Tail / Q000737:chemistry* / Q000378:metabolism; D013087:Spermatids / Q000737:chemistry / Q000378:metabolism; D013090:Spermatocytes / Q000737:chemistry / Q000378:metabolism; D013091:Spermatogenesis / Q000502:physiology; D013735:Testicular Hormones / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D013737:Testis / Q000737:chemistry* / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D015815:Cell Adhesion Molecules; C483816:Crisp2 protein, mouse; D006023:Glycoproteins; D008565:Membrane Proteins; D013735:Testicular Hormones; C474460:germ cell-specific protein 1, mouse