Phospho-Rpb1 CTD (Ser2/5) Antibody

• 产品编号：CST-4735S      品牌：CST       原厂货号：4735S
• 产品分类：抗体 > 一抗 > 磷酸化抗体
• 应用分类：

描述：

Phospho-Rpb1 CTD (Ser2/5) Antibody能够检测内源性羧基末端结构域七肽重复序列[Tyr1,Ser2,Pro3,Thr4,Ser5,Pro6,Ser7]丝氨酸（2,5位）双磷酸化和丝氨酸（2,5位）单磷酸化的Rpb1蛋白。该多克隆抗体是由合成的针对Rpb1蛋白CTD结构域七肽序列丝氨酸（2,5位）的磷酸化肽段免疫动物，采用蛋白A和多肽亲和层析技术纯化生产的。 RNA 聚合酶 II是由12种蛋白组成的复合体，参与转录、mRNA处理和转录偶联DNA修复(1,2)。最大的亚基Rpb1包含一个独特的七肽：Tyr1, Ser2, Pro3, Thr4, Ser5, Pro6, Ser7，在Rpb1的羧基末端结构域(CTD)发生高达52次的重复(3)。该结构域能够被周期蛋白依赖性激酶 (CDKs)，p44/42 MAPK，DNA-PKcs和c-Abl 磷酸化(4)。紫外线、过氧化氢和顺铂引起的DNA损伤能够导致Rpb1的泛素化和蛋白酶体降解(1,3)。激酶抑制剂5,6-二氯-1-β-D-呋喃核糖基苯并咪唑能够阻止Rpb1的泛素化表明CTD磷酸化为蛋白酶解所必需(5)。

1. Brookes, E. and Pombo, A. (2009) EMBO Rep 10, 1213-9.
2. Komarnitsky, P. et al. (2000) Genes Dev 14, 2452-60.
3. Ho, C.K. and Shuman, S. (1999) Mol Cell 3, 405-11.
4. Ng, H.H. et al. (2003) Mol Cell 11, 709-19.
5. Cheng, B. and Price, D.H. (2007) J Biol Chem 282, 21901-12.
6. Marshall, N.F. et al. (1996) J Biol Chem 271, 27176-83.
7. Krogan, N.J. et al. (2003) Mol Cell Biol 23, 4207-18.
8. Proudfoot, N.J. et al. (2002) Cell 108, 501-12.
9. Chapman, R.D. et al. (2007) Science 318, 1780-2.
10. Egloff, S. et al. (2007) Science 318, 1777-9.
11. Egloff, S. et al. (2008) Biochem Soc Trans 36, 590-4.
12. Baillat, D. et al. (2005) Cell 123, 265-76.
13. Akhtar, M.S. et al. (2009) Mol Cell 34, 387-93.
14. Egloff, S. et al. (2010) J Biol Chem 285, 20564-9.
15. Egloff, S. et al. (2012) Mol Cell 45, 111-22.

原厂资料：

Homology

Species predicted to react based on 100% sequence homology: Hamster, S. cerevisiae

Specificity / Sensitivity

Phospho-Rpb1 CTD (Ser2/5) Antibody detects endogenous levels of Rpb1 when the carboxy-termial domain heptapeptide repeat [Tyr1,Ser2,Pro3,Thr4,Ser5,Pro6,Ser7] is dually phosphorylated at Ser2 and Ser5 and singly phosphorylated at either Ser2 or Ser5.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser2/5 of the Rpb1 CTD heptapeptide. Antibodies are purified by protein A and peptide affinity chromatography.

Background

RNA polymerase II is a complex of 12 proteins that participates in transcription, mRNA processing, and transcription-coupled DNA repair (1,2). The largest subunit, Rpb1, contains a unique heptapeptide: Tyr1, Ser2, Pro3, Thr4, Ser5, Pro6, Ser7, which is repeated up to 52 times in the carboxy-terminal domain (CTD) of Rpb1 (3). This CTD is phosphorylated by cyclin-dependent kinases (CDKs), p44/42 MAPK, DNA-PKcs, and c-Abl (4). DNA damage caused by UV-light, hydrogen peroxide, or cisplatin results in ubiquitination and proteasomal degradation of Rpb1 (1,3). The kinase inhibitor 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole prevents ubiquitination of Rpb1, suggesting that CTD phosphorylation is required for proteolysis (5). 

1. Brookes, E. and Pombo, A. (2009) EMBO Rep 10, 1213-9.
2. Komarnitsky, P. et al. (2000) Genes Dev 14, 2452-60.
3. Ho, C.K. and Shuman, S. (1999) Mol Cell 3, 405-11.
4. Ng, H.H. et al. (2003) Mol Cell 11, 709-19.
5. Cheng, B. and Price, D.H. (2007) J Biol Chem 282, 21901-12.
6. Marshall, N.F. et al. (1996) J Biol Chem 271, 27176-83.
7. Krogan, N.J. et al. (2003) Mol Cell Biol 23, 4207-18.
8. Proudfoot, N.J. et al. (2002) Cell 108, 501-12.
9. Chapman, R.D. et al. (2007) Science 318, 1780-2.
10. Egloff, S. et al. (2007) Science 318, 1777-9.
11. Egloff, S. et al. (2008) Biochem Soc Trans 36, 590-4.
12. Baillat, D. et al. (2005) Cell 123, 265-76.
13. Akhtar, M.S. et al. (2009) Mol Cell 34, 387-93.
14. Egloff, S. et al. (2010) J Biol Chem 285, 20564-9.
15. Egloff, S. et al. (2012) Mol Cell 45, 111-22.

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM
NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C.
Do not aliquot the antibody.