Haploinsufficiency for the murine orthologue of Chlamydomonas PF20 disrupts spermatogenesis.
PF20 was first identified in Chlamydomonas rheinhardtii as an essential component of the axoneme central apparatus. We discovered that the mouse Pf20 gene encodes two major transcripts (2.5 and 1.4 kb), which are expressed in different patterns during spermatogenesis, yielding proteins of 71 and 35 kDa, respectively. Both proteins contain contiguous WD repeats in their C termini. The meiotically expressed 71-kDa protein is incorporated into the central apparatus, whereas the 35-kDa protein, which accumulates in postmeiotic male germ cells, is abundant in the nucleus. We disrupted the Pf20 gene domains that encode the C-terminal WD repeats in embryonic stem cells. Highly chimeric mice carrying the mutant Pf20 allele had impaired spermatogenesis with a significant loss of germ cells at the round spermatid stage, in association with disorganization of sperm axoneme structure. The mutated Pf20 allele was never transmitted, indicating that Pf20 haploinsufficiency caused the defects in spermatogenesis. The 35-kDa PF20 protein was shown to bind to meiosis-expressed gene 1 (MEIG1), a chromosome/chromatin-binding protein initially expressed during meiosis but retained in the germ cell nucleus throughout later stages of spermatogenesis. Our findings reveal an essential role for Pf20 in mouse spermatogenesis, sustaining postmeiotic germ cell viability. The different patterns of expression of the two PF20 proteins suggest the possibility that the Pf20 gene has multiple functions during spermatogenesis.
Center for Research on Reproduction and Women's Health, University of Pennsylvania Medical Center, Philadelphia, PA 19104, USA.
Zhibing
Z
Zhang
Igor
I
Kostetskii
Stuart B
SB
Moss
Brian H
BH
Jones
Clement
C
Ho
Hongbin
H
Wang
Tatsuro
T
Kishida
George L
GL
Gerton
Glenn L
GL
Radice
Jerome F
JF
Strauss
D000818:Animals; D018797:Cell Cycle Proteins; D002467:Cell Nucleus / Q000378:metabolism; D002678:Chimera / Q000235:genetics / Q000378:metabolism; D051383:GRB10 Adaptor Protein; D018390:Gene Targeting; D012739:Gonadal Steroid Hormones / Q000097:blood; D008297:Male; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D009687:Nuclear Proteins; D010750:Phosphoproteins; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D013091:Spermatogenesis / Q000235:genetics* / Q000502:physiology
D018797:Cell Cycle Proteins; D012739:Gonadal Steroid Hormones; C500770:Meig1 protein, mouse; D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins; C107014:PF20 protein, Chlamydomonas; C491467:PF20 protein, mouse; D010750:Phosphoproteins; D020033:Protein Isoforms; D011506:Proteins; D015800:Protozoan Proteins; D051383:GRB10 Adaptor Protein