共查询到20条相似文献,搜索用时 31 毫秒
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Functional coexpression of serine protein kinase SRPK1 and its substrate ASF/SF2 in Escherichia coli
Bai-Gong Yue Paul Ajuh Gran Akusjrvi Angus I. Lamond Jan-Peter Kreivi 《Nucleic acids research》2000,28(5):e14
Mammalian proteins expressed in Escherichia coli are used in a variety of applications. A major drawback in producing eukaryotic proteins in E.coli is that the bacteria lack most eukaryotic post-translational modification systems, including serine/threonine protein kinase(s). Here we show that a eukaryotic protein can be phosphorylated in E.coli by simultaneous expression of a mammalian protein kinase and its substrate. We show that in bacteria expressing SRPK1, ASF/SF2 becomes phosphorylated to a degree resembling native ASF/SF2 present in interphase HeLa cell nuclei. The E.coli phosphorylated ASF/SF2 is functional in splicing and, contrary to the unphosphorylated protein, soluble under native conditions. 相似文献
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Murata T Noda C Saito S Kawashima D Sugimoto A Isomura H Kanda T Yokoyama KK Tsurumi T 《The Journal of biological chemistry》2011,286(25):22007-22016
Reactivation of the Epstein-Barr virus from latency is dependent on expression of the BZLF1 viral immediate-early protein. The BZLF1 promoter (Zp) normally exhibits only low basal activity but is activated in response to chemical inducers such as 12-O-tetradecanoylphorbol-13-acetate and calcium ionophore. We found that Jun dimerization protein 2 (JDP2) plays a significant role in suppressing Zp activity. Reporter, EMSA, and ChIP assays of a Zp mutant virus revealed JDP2 association with Zp at the ZII cis-element, a binding site for CREB/ATF/AP-1. Suppression of Zp activity by JDP2 correlated with HDAC3 association and reduced levels of histone acetylation. Although introduction of point mutations into the ZII element of the viral genome did not increase the level of BZLF1 production, silencing of endogenous JDP2 gene expression by RNA interference increased the levels of viral early gene products and viral DNA replication. These results indicate that JDP2 plays a role as a repressor of Zp and that its replacement by CREB/ATF/AP-1 at ZII is crucial to triggering reactivation from latency to lytic replication. 相似文献
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M A Martinez-Balbás A J Bannister K Martin P Haus-Seuffert M Meisterernst T Kouzarides 《The EMBO journal》1998,17(10):2886-2893
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Activating transcription factor-2 DNA-binding activity is stimulated by phosphorylation catalyzed by p42 and p54 microtubule-associated protein kinases. 总被引:14,自引:0,他引:14
H A Abdel-Hafiz L E Heasley J M Kyriakis J Avruch D J Kroll G L Johnson J P Hoeffler 《Molecular endocrinology (Baltimore, Md.)》1992,6(12):2079-2089