Linked Life Data

14645369

Source:http://linkedlifedata.com/resource/pubmed/id/14645369

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
8
pubmed:dateCreated
2004-2-17
pubmed:abstractText
Protein kinase regulated by RNA (PKR) plays important roles in many cellular processes including virus multiplication and cell growth, differentiation, and apoptosis. The promoter of the PKR gene possesses a novel 15-bp element designated KCS, positioned upstream of a consensus interferon (IFN)-stimulated response element, that is required for both basal and interferon-inducible transcription. Protein binding to the KCS element is not dependent upon IFN treatment and correlates with transcriptional activity of the PKR promoter. The identity of KCS-binding proteins (KBP) that selectively bind at the KCS element is largely unknown, except for the transcription factor Sp1. We now have purified KBP from HeLa cell nuclear extracts by ion-exchange and DNA-affinity chromatography steps and then identified four constituent proteins of the KBP complex by mass spectrometry and immunochemistry: KBP120 and KBP45 are the damaged DNA-binding protein subunits, p127 DDB1 and p48 DDB2, respectively; KBP100 is the transcription factor Sp1; and KBP35 is the heterogeneous nuclear ribonucleoprotein A1. The steady-state levels of these four KCS-binding proteins in human cells are not altered by IFN treatment. Components of the KBP complex bind selectively and constitutively to the KCS element in the absence of IFN treatment, both in vitro as measured by competition electrophoretic mobility shift assay (EMSA) and DNA pull-down assays and in vivo as measured by chromatin immunoprecipitation assays. Depletion of DDB2 by antisense strategy reduces KBP complex formation by EMSA. These results provide new insight into the biochemical identity and activity of proteins involved in PKR promoter function.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
20
pubmed:volume
279
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
7313-21
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed-meshheading:14645369-Amino Acid Sequence, pubmed-meshheading:14645369-Blotting, Western, pubmed-meshheading:14645369-Cell Nucleus, pubmed-meshheading:14645369-Chromatin, pubmed-meshheading:14645369-Chromatography, Ion Exchange, pubmed-meshheading:14645369-DNA, pubmed-meshheading:14645369-DNA Damage, pubmed-meshheading:14645369-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:14645369-HeLa Cells, pubmed-meshheading:14645369-Heterogeneous-Nuclear Ribonucleoprotein Group A-B, pubmed-meshheading:14645369-Humans, pubmed-meshheading:14645369-Immunohistochemistry, pubmed-meshheading:14645369-Interferons, pubmed-meshheading:14645369-Mass Spectrometry, pubmed-meshheading:14645369-Models, Biological, pubmed-meshheading:14645369-Molecular Sequence Data, pubmed-meshheading:14645369-Oligonucleotides, Antisense, pubmed-meshheading:14645369-Precipitin Tests, pubmed-meshheading:14645369-Promoter Regions, Genetic, pubmed-meshheading:14645369-Protein Binding, pubmed-meshheading:14645369-Sp1 Transcription Factor, pubmed-meshheading:14645369-eIF-2 Kinase
pubmed:year
2004
pubmed:articleTitle
DNA damage-binding proteins and heterogeneous nuclear ribonucleoprotein A1 function as constitutive KCS element components of the interferon-inducible RNA-dependent protein kinase promoter.
pubmed:affiliation
Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, California 93106, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.