Zebrafish Dnd protein binds to 3'UTR of geminin mRNA and regulates its expression.
Dnd (dead end) gene encodes an RNA binding protein and is specifically expressed in primordial germ cells (PGCs) as a vertebrate-specific component of the germ plasma throughout embryogenesis. By utilizing a technique of specific nucleic acids associated with proteins (SNAAP), 13 potential target mRNAs of zebrafish Dnd (ZDnd) protein were identified from 8-cell embryo, and 8 target mRNAs have been confirmed using an RT-PCR analysis. Of the target mRNAs, the present study is focused on the regulation of geminin, which is an inhibitor of DNA replication. Using electrophoretic mobility shift assay (EMSA), we demonstrated that ZDND protein bound the 67-nucleotide region from 864 to 931 in the 3'UTR of geminin mRNA, a sequence containing 60.29% of uridine. Results from a dual-luciferase assay in HEK293 cells showed that ZDND increases the translation of geminin. Taken together, the identification of target mRNA for ZDnd will be helpful to further explore the biological function of Dnd in zebrafish germ-line development as well as in cancer cells.
Chen
Shu
S
Department of Medical Genetics, West China Hospital, Sichuan University, Chengdu, P R China.
Zeng
Mei
M
Sun
Huaqin
H
Deng
Wenqian
W
Lu
Yilu
Y
Tao
Dachang
D
Liu
Yunqiang
Y
Zhang
Sizhong
S
Ma
Yongxin
Y
D020413:3' Untranslated Regions; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D017931:DNA Primers; D024202:Electrophoretic Mobility Shift Assay; D005786:Gene Expression Regulation; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015027:Zebrafish / Q000196:embryology; D029961:Zebrafish Proteins / Q000235:genetics* / Q000378:metabolism
D020413:3' Untranslated Regions; D017931:DNA Primers; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D029961:Zebrafish Proteins; C539707:dnd1 protein, zebrafish