PES1蛋白与雄激素受体的相互作用及定位
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摘要
目的:研究PES1蛋白与雄激素受体(AR)之间的相互作用。方法:利用免疫共沉淀实验检测PES1蛋白与AR之间的相互作用,并进行相互作用定位;利用Western印迹研究PES1对乳腺癌细胞内AR表达水平的影响。结果:免疫共沉淀实验显示PES1蛋白与AR存在相互作用;PES1蛋白的1~110、111~220、221~320和311~588氨基酸残基(aa)区域均能与AR结合,415~588 aa不能结合AR;AR的651~918 aa区域与PES1结合。PES1不能调节乳腺癌细胞AR的表达水平。结论:PES1多个区域均能与AR相互作用,并且主要结合在AR的转录激活结构域2,为进一步探讨PES1对AR功能的调节奠定了基础。
Objective: To identify the interaction of PES1 and Androgen receptor(AR). Methods: Co-immuno-precipitation was performed to detect and map the interaction of PES1 and AR. The expression level of AR influ-enced by PES1 protein in breast cancer cells was investigated by Western blotting. Results: PES1 could interactwith AR. PES11-110, PES1111-220, PES1221-322, and PES1311-588 interacted with AR, whereas PES1415-588 did not. On theother hand, PES1 interacted with the amino acid region 651~918 of AR. PES1 could not regulate the expressionlevel of AR in breast cancer cells. Conclusion: Many regions of PES1 can interact with AR. PES1 interacts withthe activation function 2 domain of AR. This study lays a foundation for further study on the regulation of PES1on AR function.
Objective: To identify the interaction of PES1 and Androgen receptor(AR). Methods: Co-immuno-precipitation was performed to detect and map the interaction of PES1 and AR. The expression level of AR influ-enced by PES1 protein in breast cancer cells was investigated by Western blotting. Results: PES1 could interactwith AR. PES11-110, PES1111-220, PES1221-322, and PES1311-588 interacted with AR, whereas PES1415-588 did not. On theother hand, PES1 interacted with the amino acid region 651~918 of AR. PES1 could not regulate the expressionlevel of AR in breast cancer cells. Conclusion: Many regions of PES1 can interact with AR. PES1 interacts withthe activation function 2 domain of AR. This study lays a foundation for further study on the regulation of PES1on AR function.
引文
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[15]Holzel M M,Rohrmoser M,Schlee T,et al.MammalianWDR12 is a novel member of the Pes1-Bop1 complex andis required for ribosome biogenesis and cell proliferation[J].JCell Biol,2005,170(3):367-378.
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[17]Cheng L,Li J P,et al.PES1 promotes breast cancer by dif-ferentially regulating ERαand ERβ[J].J Clin Invest,2012,122(8):2857-2870.
[18]Li J,Yu L,Zhang H Z,et al.Down-regulation of pescadilloinhibits proliferation and tumorigenicity of breast cancer cells[J].Cancer Sci,2009,100(12):2255-2260.
[2]Brinkmann A O,Faber P W,Van Rooij H C,et al.The human androgen receptor:domain structure,genomic organization and regulation of expression[J].J Steroid Biochem,1989,34(1-6):307-310.
[3]Bevan C L,Hoare S,Claessens F,et al.The AF1 and AF2domains of the androgen receptor interact with distinct regions of SRC1[J].Mol Cell Biol,1999,19(12):8383-8392.
[4]Dehm S M,Tindall D J.Molecular regulation of androgen action in prostate cancer[J].J Cell Biochem,2006,99(2):333-344.
[5]Keren I K,Susan H,Alex G,et al.A genome-wide RNA in-terference screen identifies new regulators of androgen recep-tor function in prostate cancer cells[J].Genome Res,2013,23(4):581-591.
[6]Moe R E,Anderson B O.Androgens and androgen receptors:a clinically neglected sector in breast cancer biology[J].JSurg Oncol,2007,95(6):437-439.
[7]Higgins M J,Wolff A C.The androgen receptor in breast can-cer:learning from the past[J].Breast Cancer Res Treat,2010,124(3):619-621.
[8]Park S,Koo J S,Kim M S,et al.Androgen receptor expres-sion is significantly associated with better outcomes in estro-gen receptor-positive breast cancers[J].Ann Oncol,2011,22(8):1755-1762.
[9]Ni M,Chen Y,Lim E,et al.Targeting androgen receptor inestrogen receptor-negative breast cancer[J].Cancer Cell,2011,20(1):119-131.
[10]Du Y C,Stillman B.Yphlp,an ORC-interacting protein:po-tential links between cell proliferation control,DNA replica-tion,and ribosome biogenesis[J].Cell,2002,109(7):835-848.
[11]Holzel M,Grimm T,Rohrmoser M,et al.The BRCT domainof mammalian PES1 is crucial for nucleolar localization andrRNA processing[J].Nucleic Acids Res,2007,35(3):789-800.
[12]Kinoshita Y,Jarell A D,Flaman J M,et al.Pescadillo,anovel cell cycleregulatory protein abnormally expressed in ma-lignant cells[J].J Biol Chem,2001,276(9):6656-6665.
[13]Lerch-Gaggl A,Haque J,Li J X,et al.Pescadillo is essen-tial for nucleolar assembly,ribosome biogenesis,and mammali-an cell proli-feration[J].J Biol Chem,2002,277(47):45347-45355.
[14]Prisco M,Maiorana A,Guerzoni C,et al.Role of pescadilloand upstream binding factor in the proliferation and differenti-ation of murine myeloid Cells[J].Mol Cell Biol,2004,24(12):5421-5433.
[15]Holzel M M,Rohrmoser M,Schlee T,et al.MammalianWDR12 is a novel member of the Pes1-Bop1 complex andis required for ribosome biogenesis and cell proliferation[J].JCell Biol,2005,170(3):367-378.
[16]Zhang H,Fang Y,Huang C,et al.Human pescadillo induceslarge-scale chromatin unfolding[J].Sci China C Life Sci,2005,48(3):270-276.
[17]Cheng L,Li J P,et al.PES1 promotes breast cancer by dif-ferentially regulating ERαand ERβ[J].J Clin Invest,2012,122(8):2857-2870.
[18]Li J,Yu L,Zhang H Z,et al.Down-regulation of pescadilloinhibits proliferation and tumorigenicity of breast cancer cells[J].Cancer Sci,2009,100(12):2255-2260.