The protein encoded by the germ plasm RNA Germes associates with dynein light chains and functions in Xenopus germline development.
Germ plasm plays a prominent role in germline formation in a large number of animal taxons. We previously identified a novel maternal RNA named Germes associated with Xenopus germ plasm. In the present work, we addressed possible involvement of Germes protein in germ plasm function. Expression in oocytes followed by confocal microscopy revealed that the EGFP fused to Germes, in contrast to the free EGFP, co-localized with the germ plasm. Overexpression of intact Germes and Germes lacking both leucine zipper motifs (GermesDeltaLZs) resulted in a statistically significant reduction of the number of primordial germ cells (PGCs). Furthermore, the GermesDeltaLZs mutant inhibited PGC migration and produced abnormalities in germ plasm intra-cellular distribution at tailbud stages. To begin unraveling biochemical interactions of Germes during embryogenesis, we searched for Germes partners using yeast two-hybrid (YTH) system. Two closely related sequences were identified, encoding Xenopus dynein light chains dlc8a and dlc8b. Tagged versions of Germes and dlc8s co-localize in VERO cells upon transient expression and can be co-immunoprecipitated after injection of the corresponding RNAs in Xenopus embryos, indicating that their interactions occur in vivo. We conclude that Germes is involved in organization and functioning of germ plasm in Xenopus, probably through interaction with motor complexes.
Berekelya
Lyubov A
LA
Engelhardt Institute of Molecular Biology, Moscow, Russia.
Mikryukov
Alexander A
AA
Luchinskaya
Natalia N
NN
Ponomarev
Maxim B
MB
Woodland
Hugh R
HR
Belyavsky
Alexander V
AV
D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D049109:Cell Proliferation; D002478:Cells, Cultured; D003001:Cloning, Molecular; D029721:Drosophila Proteins / Q000378:metabolism*; D004398:Dyneins; D005260:Female; D005456:Fluorescent Dyes / Q000032:analysis; D005854:Germ Cells / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000032:analysis; D009154:Mutation; D009865:Oocytes / Q000254:growth & development / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000502:physiology*; D015854:Up-Regulation; D014981:Xenopus / Q000196:embryology* / Q000254:growth & development; D029867:Xenopus Proteins / Q000235:genetics / Q000502:physiology*
D002352:Carrier Proteins; D029721:Drosophila Proteins; D005456:Fluorescent Dyes; C525160:Germes protein, Xenopus; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D029867:Xenopus Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D004398:Dyneins