A conserved zinc finger transcription factor GATA involving in the hemocyte production of scallop Chlamys farreri.
GATA are a family of transcription factors characterized by their ability to bind to the DNA sequence "GATA", and involved in a myriad of cellular processes. GATA1/2/3 factors are known as the hematopoietic GATA factors, which play dominated roles in regulating hematopoiesis. In the present study, a gene encoding GATA transcription factor (designed as CfGATA) was cloned and characterized from the scallop Chlamys farreri. The full-length cDNA of CfGATA is of 2058 bp encoding a predicted polypeptide of 457 amino acids with two conserved zinc finger domains, which shared high similarity with other reported GATA1/2/3 proteins. The mRNA transcripts of CfGATA showed higher expression in gills, hepatopancreas, hemocytes and heart, and the CfGATA protein expressed in HEK293 cells was found to be localized specifically in the nuclei. The recombinant CfGATA protein (rCfGATA) exhibited strong ability to bind specific WGATAR DNA sequence by electrophoretic mobility shift assay in vitro. After CfGATA gene was silenced by RNA interference, the hemocyte renewal rate and circulating total hemocyte count (THC) decreased significantly, which was 7.85-fold and 19.46-fold lower than that of PBS control, respectively (P < 0.05). After LPS stimulation, the expression level of CfGATA mRNA decreased significantly in the hemocytes of PBS or EGFP dsRNA treated scallops, which was accompanied by the increase of hemocyte renewal rate and the reduced circulating THC at 24 h. In contrast, the hemocyte renewal rate and circulating THC did not change significantly in CfGATA gene interfered scallops after LPS stimulation. These results suggested that CfGATA, as a conserved GATA1/2/3 transcription factor, plays essential roles in regulating hemocyte production of scallop.
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Rd., Qingdao, Shandong 266071, China; University of Chinese Academy of Sciences, Beijing 100049, China.
Feng
F
Yue
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Rd., Qingdao, Shandong 266071, China.
Zhi
Z
Zhou
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Rd., Qingdao, Shandong 266071, China.
Lingling
L
Wang
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Rd., Qingdao, Shandong 266071, China.
Mengqiang
M
Wang
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Rd., Qingdao, Shandong 266071, China. Electronic address: lshsong@ms.qdio.ac.cn.
Linsheng
L
Song
D000704:Analysis of Variance; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D005455:Fluorescent Antibody Technique; D050980:GATA Transcription Factors / Q000235:genetics* / Q000378:metabolism; D006434:Hemocytes / Q000502:physiology*; D008969:Molecular Sequence Data; D049897:Pectinidae / Q000166:cytology / Q000235:genetics*; D034622:RNA Interference; D011994:Recombinant Proteins / Q000378:metabolism; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D050980:GATA Transcription Factors; D011994:Recombinant Proteins; D004247:DNA
Chlamys farreri; GATA; Hematopoiesis; Hemocyte production; Transcription factor