TY - JOUR
T1 - Transcripts immunoprecipitated with Sxl protein in primordial germ cells of Drosophila embryos
AU - Ota, Ryoma
AU - Morita, Shumpei
AU - Sato, Masanao
AU - Shigenobu, Shuji
AU - Hayashi, Makoto
AU - Kobayashi, Satoru
N1 - Funding Information:
also thank the Bloomington Drosophila Stock Center and Vienna Drosophila Resource Center for providing us with fly stocks, and the Developmental Studies Hybridoma Bank for antibodies. This work was supported in part by Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (JSPS) (KAKENHI Grant Numbers: 25114002 and 24247011).
Publisher Copyright:
© 2017 Japanese Society of Developmental Biologists
PY - 2017/12
Y1 - 2017/12
N2 - In Drosophila, Sex lethal (Sxl), an RNA binding protein, is required for induction of female sexual identity in both somatic and germline cells. Although the Sxl-dependent feminizing pathway in the soma was previously elucidated, the downstream targets for Sxl in the germline remained elusive. To identify these target genes, we selected transcripts associated with Sxl in primordial germ cells (PGCs) of embryos using RNA immunoprecipitation coupled to sequencing (RIP-seq) analysis. A total of 308 transcripts encoded by 282 genes were obtained. Seven of these genes, expressed at higher levels in PGCs as determined by microarray and in situ hybridization analyses, were subjected to RNAi-mediated functional analyses. Knockdown of Neos, Kap-alpha3, and CG32075 throughout germline development caused gonadal dysgenesis in a sex-dependent manner, and Su(var)2-10 knockdown caused gonadal dysgenesis in both sexes. Moreover, as with knockdown of Sxl, knockdown of Su(var)2-10 in PGCs gave rise to a tumorous phenotype of germline cells in ovaries. Because this phenotype indicates loss of female identity of germline cells, we consider Su(var)2-10 to be a strong candidate target of Sxl in PGCs. Our results represent a first step toward elucidating the Sxl-dependent feminizing pathway in the germline.
AB - In Drosophila, Sex lethal (Sxl), an RNA binding protein, is required for induction of female sexual identity in both somatic and germline cells. Although the Sxl-dependent feminizing pathway in the soma was previously elucidated, the downstream targets for Sxl in the germline remained elusive. To identify these target genes, we selected transcripts associated with Sxl in primordial germ cells (PGCs) of embryos using RNA immunoprecipitation coupled to sequencing (RIP-seq) analysis. A total of 308 transcripts encoded by 282 genes were obtained. Seven of these genes, expressed at higher levels in PGCs as determined by microarray and in situ hybridization analyses, were subjected to RNAi-mediated functional analyses. Knockdown of Neos, Kap-alpha3, and CG32075 throughout germline development caused gonadal dysgenesis in a sex-dependent manner, and Su(var)2-10 knockdown caused gonadal dysgenesis in both sexes. Moreover, as with knockdown of Sxl, knockdown of Su(var)2-10 in PGCs gave rise to a tumorous phenotype of germline cells in ovaries. Because this phenotype indicates loss of female identity of germline cells, we consider Su(var)2-10 to be a strong candidate target of Sxl in PGCs. Our results represent a first step toward elucidating the Sxl-dependent feminizing pathway in the germline.
KW - germline
KW - RNA immunoprecipitation and RNA sequencing
KW - sex determination
KW - Su(var)2-10
KW - Sxl
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U2 - 10.1111/dgd.12408
DO - 10.1111/dgd.12408
M3 - Article
C2 - 29124738
AN - SCOPUS:85033503500
SN - 0012-1592
VL - 59
SP - 713
EP - 723
JO - Development Growth and Differentiation
JF - Development Growth and Differentiation
IS - 9
ER -