SUMO Modification of NZFP Mediates Transcriptional Repression through TBP Binding

다국어 초록 (Multilingual Abstract)

The negatively regulating zinc finger protein (NZFP) is an essential transcription repressor required for early devel-opment during gastrulation in Xenopus laevis. In this study, we found that NZFP interacts with the small ubiq-uitin-like modifier (SUMO) conjugation E2 enzyme, Ubc9, and contains three putative SUMO conjugation sites. Studies with NZFP mutants containing mutations at the putative SUMO conjugation sites showed that these sites were able to be modified independently with SUMO. NZFP was found to be localized in the same nuclear bodies with SUMO-1. However, sumoylation of NZFP did not play a role either in the translocation of NZFP into the nucleus or on nuclear body formation. While wild type NZFP showed significant transcriptional repression, SUMO-conjugation site mutants manifested a decrease in transcriptional repression activity which is reversely proportional to the amount of sumoylation. The sumoylation defective mutant lost its TBP binding activity, while wild type NZFP interacted with TBP and inhibited transcription complex formation. These results strongly suggest that the sumoylation of NZFP facilitates NZFP to bind to TBP and the NZFP/TBP complex then represses the transcription of the target gene by in-hibiting basal transcription complex formation.

참고문헌 (Reference)

1 Zheng, G, "ZNF76, a novel transcriptional repressor targeting TATA-binding protein, is modulated by sumoylation" 279 : 42410-42421, 2004

2 Kim, K.I, "Versatile protein tag, SUMO: Its enzymology and biological function" 191 : 257-268, 2002

3 Kim, J., "Transcriptional activity of CCAAT/enhancer-binding proteins is controlled by a conserved inhibitory domain that is a target for sumoylation" 277 : 38037-38044, 2002

4 Sapetschnig, A., "Transcription factor Sp3 is silenced through SUMO modification by PIAS1" 21 : 5206-5215, 2002

5 Buscarlet, M, "The ‘Marx’ of Groucho on development and disease" 17 : 353-361, 2007

6 Li, S.J, "The yeast gene ULP2 (SMT4) gene encodes a novel protease specific for the ubiquitin-like Smt3 protein" 20 : 2367-2377, 2000

7 Gareau, J.R, "The SUMO pathway: emerging mechanisms that shape specificity, conjugation and recognition" 11 : 861-871, 2010

8 Kovall, R.A., "Structures of CSL, Notch and Mastermind proteins: piecing together an active transcription complex" 17 : 117-127, 2007

9 Gill, G, "Something about SUMO inhibits transcription" 15 : 536-541, 2005

10 Tian, S, "Small ubiquitin-related modifier-1 (SUMO-1) modification of the glucocorticoid receptor" 367 : 907-911, 2002

11 Ross, S., "SUMO-1 modification represses Sp3 transcriptional activation and modulates its subnuclear localization" 10 : 831-842, 2002

12 Rodriguez, M.S, "SUMO-1 conjugation in vivo requires both a consensus modification motif and nuclear targeting" 276 : 12654-12659, 2001

13 Yang, S.H, "SUMO promotes HDACmediated transcriptional repression" 13 : 611-617, 2004

14 Gill, G., "SUMO and ubiquitin in the nucleus: different functions, similar mechanisms" 18 : 2046-2059, 2004

15 Zhong, S, "Role of SUMO-1-modified PML in nuclear body formation" 95 : 2748-2752, 2000

16 Liu, B, "Regulation of the sumoylation system in gene expression" 20 : 288-293, 2008

17 Hong, Y, "Regulation of heat shock transcription factor 1 by stress-induced SUMO-1 modification" 276 : 40263-40267, 2001

18 Gong, L, "Preferential interaction of sentrin with a ubiquitin-conjugating enzyme, Ubc9" 272 : 28198-28201, 1997

19 Le Drean, Y, "Potentiation of glucocorticoid receptor transcriptional activity by sumoylation" 143 : 3482-3489, 2002

20 Schmidt, D, "PIAS/SUMO: new partners in transcriptional regulation" 60 : 2561-2574, 2003

21 Fernandez-Martinez, J, "Overlap of nuclear localisation signal and specific DNA-binding residues within the zinc finger domain of PacC" 334 : 667-684, 2003

22 Knöchel, W, "Evolutionary conserved modules associated with zinc fingers in Xenopus laevis" 86 : 6097-6100, 1989

23 Arce, L, "Diversity of LEF/TCF action in development and disease" 25 : 7492-7504, 2006

24 Chang, C.C, "Daxx mediates the small ubiquitin-like modifier-dependent transcriptional repression of Smad4" 280 : 10164-10173, 2005

25 Kim, Y.H, "Covalent modification of the homeodomain-interacting protein kinase 2 (HIPK2) by the ubiquitin-like protein SUMO-1" 96 : 12350-12355, 1999

26 Poukka, H, "Covalent modification of the androgen receptor by small ubiquitin- like modifier 1 (SUMO-1)" 97 : 14145-14150, 2000

27 Bies, J, "Covalent attachment of the SUMO-1 protein to the negative regulatory domain of the c- Myb transcription factor modifies its stability and transactivation capacity" 277 : 8999-9009, 2002

28 Müller, S, "Conjugation with the ubiquitin-related modifier SUMO-1 regulates the partitioning of PML within the nucleus" 17 : 61-70, 1998

29 Geiss-Friedlander R, "Concepts in sumoylation: a decade on" 8 : 947-956, 2007

30 Kim, M, "A novel TBP-interacting zinc finger protein represses transcription by inhibiting the recruitment of TFIIA and TFIIB" 306 : 231-238, 2003

31 Kim, M., "A novel TBP-interacting zinc finger protein functions in early development of Xenopus laevis" ACADEMIC PRESS INC ELSEVIER SCIENCE 306 (306): 0000001106-0000001111, 2003