Molecular characterization of tripartite motif protein 25 (TRIM25) involved in ERα-mediated transcription in the Korean rose bitterling Rhodeus uyekii.
Tripartite motif-containing 25 (TRIM25), also known as estrogen-responsive finger protein (EFP), plays an essential role in cell proliferation and innate immunity. In the present study, we isolated and characterized the TRIM25 cDNA of the Korean rose bitterling Rhodeus uyekii, designated RuTRIM25. It encodes an open reading frame of 669 amino acids containing an N-terminal RBCC motif composed of a RING domain, two B boxes, and a coiled-coil domain and a C-terminal B30.2 (PRY/SPRY) domain. RuTRIM25 shows strong homology (79.7%) to zebrafish TRIM25 and shared 32.4-28.8% homology with TRIM25 from other species, including mammals. RuTRIM25 mRNA was expressed ubiquitously. It was highly expressed in the ovary, spleen, and liver and moderately in the stomach and intestine of normal Korean rose bitterling. The intracellular localization of RuTRIM25 in HEK293T cells was diffusely localized in the cytoplasm and its RING domain deletion mutant (RuTRIM25ΔR) was detected diffusely with some aggregates in the cytoplasm. RuTRIM25, but not RuTRIM25ΔR, is ubiquitinated in vivo. Ectopic expression of RuTRIM25 synergistically activated the estrogen receptor (ER)-mediated luciferase reporter activity in a dose-dependent manner in HEK293T cells. Together, these results suggest that the RuTRIM25 regulates the ER-mediated transcription in fish similarly to its mammalian counterpart.
Biotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, Republic of Korea. konghj@nfrdi.go.kr
Hee Jeong
HJ
Kong
Ye Ji
YJ
Lee
Jihye
J
Shin
Hyun Kook
HK
Cho
Woo-Jin
WJ
Kim
Hyung Soo
HS
Kim
Jaehun
J
Cheong
Young Chang
YC
Sohn
Sang-Jun
SJ
Lee
Bong-Seok
BS
Kim
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003530:Cyprinidae; D004591:Electrophoresis, Polyacrylamide Gel; D047628:Estrogen Receptor alpha / Q000378:metabolism*; D029941:Fish Proteins / Q000737:chemistry / Q000145:classification / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000378:metabolism*
D047628:Estrogen Receptor alpha; D029941:Fish Proteins; D044767:Ubiquitin-Protein Ligases