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禽类淋巴细胞中C/EBP相关复合物的鉴定。

Identification of a C/EBP-Rel complex in avian lymphoid cells.

作者信息

Diehl J A, Hannink M

机构信息

Biochemistry Department, University of Missouri, Columbia 65212.

出版信息

Mol Cell Biol. 1994 Oct;14(10):6635-46. doi: 10.1128/mcb.14.10.6635-6646.1994.

DOI:10.1128/mcb.14.10.6635-6646.1994
PMID:7935382
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC359193/
Abstract

Protein-protein interactions between the CCAAT box enhancer-binding proteins (C/EBP) and the Rel family of transcription factors have been implicated in the regulation of cytokine gene expression. We have used sequence-specific DNA affinity chromatography to purify a complex from avian T cells that binds to a consensus C/EBP motif. Our results provide evidence that Rel-related proteins are components of the C/EBP-DNA complex as a result of protein-protein interactions with the C/EBP proteins. A polyclonal antiserum raised against the Rel homology domain of v-Rel and antisera raised against two human RelA-derived peptides specifically induced a supershift of the C/EBP-DNA complex in mobility shift assays using the affinity-purified C/EBP. In addition, several kappa B-binding proteins copurified with the avian C/EBP complex through two rounds of sequence-specific DNA affinity chromatography. The kappa B-binding proteins are distinct from the C/EBP proteins that directly contact DNA containing the C/EBP binding site. The identification of a protein complex that binds specifically to a consensus C/EBP site and contains both C/EBP and Rel family members suggests a novel mechanism for regulation of gene expression by Rel family proteins.

摘要

CCAAT盒增强子结合蛋白（C/EBP）与Rel转录因子家族之间的蛋白质-蛋白质相互作用与细胞因子基因表达的调控有关。我们利用序列特异性DNA亲和层析从禽类T细胞中纯化出一种与共有C/EBP基序结合的复合物。我们的结果提供了证据，表明Rel相关蛋白是C/EBP-DNA复合物的组成成分，这是由于其与C/EBP蛋白发生蛋白质-蛋白质相互作用的结果。针对v-Rel的Rel同源结构域产生的多克隆抗血清以及针对两种源自人RelA的肽产生的抗血清，在使用亲和纯化的C/EBP进行的迁移率变动分析中，特异性地诱导了C/EBP-DNA复合物的超迁移。此外，通过两轮序列特异性DNA亲和层析，几种κB结合蛋白与禽类C/EBP复合物共同纯化。κB结合蛋白不同于直接接触含有C/EBP结合位点的DNA的C/EBP蛋白。鉴定出一种特异性结合共有C/EBP位点且同时包含C/EBP和Rel家族成员的蛋白质复合物，提示了Rel家族蛋白调控基因表达的一种新机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/a50d9a672e28/molcellb00010-0247-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/d8df3509726e/molcellb00010-0242-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/2126c4658f57/molcellb00010-0243-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/34b64885a986/molcellb00010-0243-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/1e47484c82fb/molcellb00010-0244-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/5dcc9fb6c443/molcellb00010-0244-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/110c071e5cd4/molcellb00010-0245-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/fe99998821d0/molcellb00010-0246-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/03958b8e3f7c/molcellb00010-0246-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/4796b31958ef/molcellb00010-0247-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/a50d9a672e28/molcellb00010-0247-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/d8df3509726e/molcellb00010-0242-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/2126c4658f57/molcellb00010-0243-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/34b64885a986/molcellb00010-0243-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/1e47484c82fb/molcellb00010-0244-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/5dcc9fb6c443/molcellb00010-0244-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/110c071e5cd4/molcellb00010-0245-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/fe99998821d0/molcellb00010-0246-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/03958b8e3f7c/molcellb00010-0246-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/4796b31958ef/molcellb00010-0247-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c91e/359193/a50d9a672e28/molcellb00010-0247-b.jpg

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