Linked Life Data

19454241

Source:http://linkedlifedata.com/resource/pubmed/id/19454241

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2009-5-22
pubmed:abstractText
To estimate the transcriptional activity of p53 in individual living mammalian cells, we constructed the enhanced green fluorescent protein-red fluorescent protein (EGFP-DsRed) reporter system with the EGFP-p53 expression vector and the reporter plasmid, which carried a p53-dependent promoter. The expression level and transcriptional activity of EGFP-p53 were determined simultaneously by green and red fluorescence signals, respectively. In this system, we could target only the cells expressing p53 at endogenous levels, as observed in UV- or adriamycin-stimulated A549 cells. Using this system, we investigated the transcriptional activity of mutant p53s in tetramerization domain. Transcriptional activities were nearly abolished by seven mutations and significantly reduced in several mutant p53s. However, under overexpression conditions, the latter mutant p53s showed activity similar to that observed in wild-type p53. These results indicated the importance of physiological concentration for p53 proteins in cells so as to analyze their activities. Fluorescence intensity distribution analysis indicated that the mutant p53s lacking transcriptional activity presented as monomer forms in the cellular extract. In most of the mutant p53s, the decrease in transcriptional activity correlated with an increase in the fraction of monomers. This reporter system can be used for estimating the transcriptional activity of mutant p53s without contribution of the cells overexpressing p53.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
1096-0309
pubmed:author
pubmed:issnType
Electronic
pubmed:day
15
pubmed:volume
387
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
249-56
pubmed:meshHeading
pubmed:year
2009
pubmed:articleTitle
Evaluation of transcriptional activity of p53 in individual living mammalian cells.
pubmed:affiliation
Laboratory of Biological Chemistry, Department of Chemistry, Faculty of Science, Hokkaido University, Sapporo 060-0810, Japan. toshi@sci.hokudai.ac.jp
pubmed:publicationType
Journal Article