Combinatorial RNA interference indicates GLH-4 can compensate for GLH-1; these two P granule components are critical for fertility in C. elegans.
We report that four putative germline RNA helicases, GLHs, are components of the germline-specific P granules in Caenorhabditis elegans. GLH-3 and GLH-4, newly discovered, belong to a multi-gene glh family. Although GLHs are homologous to Drosophila VASA, a polar granule component necessary for oogenesis and embryonic pattern formation, the GLHs are distinguished by containing multiple CCHC zinc fingers. RNA-mediated interference (RNAi) reveals the GLHs are critical for oogenesis. By RNAi at 20 degrees C, when either loss of GLH-1 or GLH-4 alone has no effect, loss of both GLH-1 and GLH-4 results in 97% sterility in the glh-1/4(RNAi) offspring of injected hermaphrodites. glh-1/4(RNAi) germlines are under-proliferated and are without oocytes. glh-1/4(RNAi) animals produce sperm; however, spermatogenesis is delayed and the sperm are defective. P granules are still present in glh-1/4(RNAi) sterile worms as revealed with antibodies against the remaining GLH-2 and GLH-3 proteins, indicating the GLHs function independently in P granule assembly. These studies reveal that C.elegans can use GLH-1 or GLH-4 to promote germline development.
Kuznicki
K A
KA
Department of Molecular Microbiology and Immunology, University of Missouri, Columbia, MO 65212, USA.
Smith
P A
PA
Leung-Chiu
W M
WM
Estevez
A O
AO
Scott
H C
HC
Bennett
K L
KL
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D017173:Caenorhabditis elegans / Q000196:embryology / Q000201:enzymology* / Q000502:physiology*; D002455:Cell Division / Q000235:genetics / Q000502:physiology; D002467:Cell Nucleus / Q000378:metabolism; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008297:Male; D008969:Molecular Sequence Data; D005810:Multigene Family; D009866:Oogenesis / Q000235:genetics / Q000502:physiology*; D010641:Phenotype; D012313:RNA / Q000378:metabolism*; D020365:RNA Helicases / Q000235:genetics / Q000502:physiology*; D017386:Sequence Homology, Amino Acid; D013091:Spermatogenesis / Q000235:genetics / Q000502:physiology*; D013696:Temperature; D016335:Zinc Fingers
D012313:RNA; D020365:RNA Helicases