Date post: | 22-Dec-2015 |
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Lab Introduction
Lab 429, Institute of Molecular Biology, Academia Sinica
PI: Bon-chu Chung, Ph.D.
Research interests
I. Steroid Function and Regulation
II. Zebrafish Development
Schematic overview of the biosynthetic pathways for steroid hormones
SF1 regulates the expression of P450 steroid hydroxylases.
Steroidogenic factor 1 (SF-1)
The Ftz-F1 box of SF-1 contains NLS sequence. (Li et al., 1999)
SF-1 is an orphan nuclear receptor.SF-1 was always found in nucleus in previous stu
dies.
SF-1
SUMO-1
SUMO-1 is a 101 amino acid polypeptide and only 1-97 a.a. can be attached to target proteins.
SF-1 has two sumoylation sites, K-119 and K-194.SUMO-1 can trigger the formation of PML nuclear
bodies.
SF-1
Cloning of FLAG-RFP-SUMO-1 fusion protein
SUMO-1 WT: 94 95 96 97CAA ACG GGG GGT Gly Gly
SUMO-1 AA (mutant): 94 95 96 97CAA ACG GCG GCT Ala Ala
Sumoylation regulates nuclear distribution of SF-1
RFP-SUMO-1 WT SF-1-GFP Merge
RFP-SUMO-1 AA SF-1-GFP Merge
WT
AA
LMB does not affect cytoplasmic distribution of SF-1
15min 45min 1h15min 1h45min
2h15min 2h45min 3h15min 3h45min
4h15min 4h45min 5h15min 5h45min
6h15min 6h45min (After LMB treatment)
SF-1 contains LMB- insensitive NES homologue SF-1
FF1bAR
SF-1
FF1b
AR
(743-817)
(743-817)
(306-381)
(306-381)
(359-436)
(359-436)
AR
AR
MRER
MRER
(743-817)
(743-817)(809-883)(386-455)
(809-883)(386-455)
(Saporita et al., 2003)
Future work
Examine the interaction of SF-1 and SUMO-1 in living cell under different stimulus.
Examine the candidate NES fragment of SF-1.
Use electro-microscopy to find out the exact location of cytoplasmic SF-1.