NF-kappa B Protein Purification from Bovine Spleen: Nucleotide Stimulation and Binding Site Specificity

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1988 Dec 1

PMID 2848241

Citations 53

Authors

M J Lenardo

A Kuang

A Gifford

D Baltimore

Affiliations

Soon will be listed here.

Abstract

The activity of the enhancer for the kappa immunoglobulin light chain gene critically depends on the presence in the nucleus of the NF-kappa B protein. We purified NF-kappa B over 50,000-fold and identified two protein species, 42 and 44 kDa, that could be eluted and renatured from a sodium dodecyl sulfate/polyacrylamide gel to give specific DNA-binding activity. Binding of the purified bovine NF-kappa B as well as that from human and murine B- or T-lymphoid cell extracts was dramatically stimulated by nucleoside triphosphates. This effect distinguished NF-kappa B from a related factor, H2-TF1. Purified NF-kappa B interacted efficiently with regulatory sequences that function during either B- or T-lymphocyte activation, including the human immunodeficiency virus enhancer and a NF-kappa B binding site we detected in the interleukin 2 enhancer.

Citing Articles

Molecular Mechanisms and Genetics of Oxidative Stress in Alzheimer's Disease.

Cioffi F, Adam R, Broersen K J Alzheimers Dis. 2019; 72(4):981-1017.

PMID: 31744008 PMC: 6971833. DOI: 10.3233/JAD-190863.

Cross-talk between NADPH oxidase-PKCα-p(38)MAPK and NF-κB-MT1MMP in activating proMMP-2 by ET-1 in pulmonary artery smooth muscle cells.

Sarkar J, Chowdhury A, Chakraborti T, Chakraborti S Mol Cell Biochem. 2016; 415(1-2):13-28.

PMID: 26910780 DOI: 10.1007/s11010-016-2673-6.

Activation of proMMP-2 by U46619 occurs via involvement of p(38)MAPK-NFκB-MT1MMP signaling pathway in pulmonary artery smooth muscle cells.

Chowdhury A, Roy S, Chakraborti T, Dey K, Chakraborti S Mol Cell Biochem. 2013; 385(1-2):53-68.

PMID: 24065388 DOI: 10.1007/s11010-013-1814-4.

Activation of RelA homodimers by tumour necrosis factor alpha: a possible transcriptional activator in human vascular endothelial cells.

Marui N, Medford R, Ahmad M Biochem J. 2005; 390(Pt 1):317-24.

PMID: 15876188 PMC: 1184585. DOI: 10.1042/BJ20041659.

Regulation of c-Rel nuclear localization by binding of Ca2+/calmodulin.

Antonsson A, Hughes K, Edin S, Grundstrom T Mol Cell Biol. 2003; 23(4):1418-27.

PMID: 12556500 PMC: 141150. DOI: 10.1128/MCB.23.4.1418-1427.2003.

References

Bradford M . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54. DOI: 10.1016/0003-2697(76)90527-3. View

Wu F, Garcia J, Harrich D, Gaynor R . Purification of the human immunodeficiency virus type 1 enhancer and TAR binding proteins EBP-1 and UBP-1. EMBO J. 1988; 7(7):2117-30. PMC: 454507. DOI: 10.1002/j.1460-2075.1988.tb03051.x. View

Wray W, Boulikas T, Wray V, Hancock R . Silver staining of proteins in polyacrylamide gels. Anal Biochem. 1981; 118(1):197-203. DOI: 10.1016/0003-2697(81)90179-2. View

Dignam J, Lebovitz R, Roeder R . Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983; 11(5):1475-89. PMC: 325809. DOI: 10.1093/nar/11.5.1475. View

Fujita T, Takaoka C, Matsui H, Taniguchi T . Structure of the human interleukin 2 gene. Proc Natl Acad Sci U S A. 1983; 80(24):7437-41. PMC: 389966. DOI: 10.1073/pnas.80.24.7437. View

Fuse A, Fujita T, Yasumitsu H, Kashima N, Hasegawa K, Taniguchi T . Organization and structure of the mouse interleukin-2 gene. Nucleic Acids Res. 1984; 12(24):9323-31. PMC: 320464. DOI: 10.1093/nar/12.24.9323. View

Jones K, Yamamoto K, Tjian R . Two distinct transcription factors bind to the HSV thymidine kinase promoter in vitro. Cell. 1985; 42(2):559-72. DOI: 10.1016/0092-8674(85)90113-8. View

Alt F, Blackwell T, DePinho R, Reth M, Yancopoulos G . Regulation of genome rearrangement events during lymphocyte differentiation. Immunol Rev. 1986; 89:5-30. DOI: 10.1111/j.1600-065x.1986.tb01470.x. View

Kadonaga J, Tjian R . Affinity purification of sequence-specific DNA binding proteins. Proc Natl Acad Sci U S A. 1986; 83(16):5889-93. PMC: 386402. DOI: 10.1073/pnas.83.16.5889. View

10.

Sen R, Baltimore D . Multiple nuclear factors interact with the immunoglobulin enhancer sequences. Cell. 1986; 46(5):705-16. DOI: 10.1016/0092-8674(86)90346-6. View

11.

Greene W, Depper J, Kronke M, Leonard W . The human interleukin-2 receptor: analysis of structure and function. Immunol Rev. 1986; 92:29-48. DOI: 10.1111/j.1600-065x.1986.tb01492.x. View

12.

Arndt K, Fink G . GCN4 protein, a positive transcription factor in yeast, binds general control promoters at all 5' TGACTC 3' sequences. Proc Natl Acad Sci U S A. 1986; 83(22):8516-20. PMC: 386961. DOI: 10.1073/pnas.83.22.8516. View

13.

Sen R, Baltimore D . Inducibility of kappa immunoglobulin enhancer-binding protein Nf-kappa B by a posttranslational mechanism. Cell. 1986; 47(6):921-8. DOI: 10.1016/0092-8674(86)90807-x. View

14.

Atchison M, PERRY R . The role of the kappa enhancer and its binding factor NF-kappa B in the developmental regulation of kappa gene transcription. Cell. 1987; 48(1):121-8. DOI: 10.1016/0092-8674(87)90362-x. View

15.

Muesing M, Smith D, Capon D . Regulation of mRNA accumulation by a human immunodeficiency virus trans-activator protein. Cell. 1987; 48(4):691-701. DOI: 10.1016/0092-8674(87)90247-9. View

16.

CROSS S, Feinberg M, Wolf J, Holbrook N, Wong-Staal F, Leonard W . Regulation of the human interleukin-2 receptor alpha chain promoter: activation of a nonfunctional promoter by the transactivator gene of HTLV-I. Cell. 1987; 49(1):47-56. DOI: 10.1016/0092-8674(87)90754-9. View

17.

Baldwin Jr A, Sharp P . Binding of a nuclear factor to a regulatory sequence in the promoter of the mouse H-2Kb class I major histocompatibility gene. Mol Cell Biol. 1987; 7(1):305-13. PMC: 365070. DOI: 10.1128/mcb.7.1.305-313.1987. View

18.

Nabel G, Baltimore D . An inducible transcription factor activates expression of human immunodeficiency virus in T cells. Nature. 1987; 326(6114):711-3. DOI: 10.1038/326711a0. View

19.

Lenardo M, PIERCE J, Baltimore D . Protein-binding sites in Ig gene enhancers determine transcriptional activity and inducibility. Science. 1987; 236(4808):1573-7. DOI: 10.1126/science.3109035. View

20.

Wu F, Garcia J, Mitsuyasu R, Gaynor R . Alterations in binding characteristics of the human immunodeficiency virus enhancer factor. J Virol. 1988; 62(1):218-25. PMC: 250522. DOI: 10.1128/JVI.62.1.218-225.1988. View