Linked Life Data

8400240

Source:http://linkedlifedata.com/resource/pubmed/id/8400240

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
6
pubmed:dateCreated
1993-10-25
pubmed:abstractText
The use of recombinant adeno-associated virus (rAAV) vectors provides a new strategy to investigate the role of specific regulatory elements and trans-acting factors in globin gene expression. We linked hypersensitivity site 2 (HS2) from the locus control region (LCR) to a A gamma-globin gene (A gamma*) mutationally marked to allow its transcript to be distinguished from endogenous gamma-globin mRNA. The vector also contains the phosphotransferase gene that confers resistance to neomycin (NeoR). HS2 region mutations within the NF-E2 motifs prevented NF-E2 binding while preserving AP-1 binding. Another set in the GATA-1 motif prevented binding of the factor. Several NeoR K562 clones containing a single unrearranged RAAV genome with the A gamma* gene linked to the native HS2 core fragment (WT), mutant NF-E2 HS2 (mut-NFE2), mutant GATA-1 HS2 (mut-GATA1), or no HS [(-)HS] were identified. In uninduced K562 cells, mut-NFE2 clones expressed A gamma* mRNA at the same level as the WT clones, compared with a lack of A gamma* signal in the (-)HS2 clones. However, hemin induction of mut-NFE2 clones did not result in an increase in the A gamma* signal above the level seen in uninduced cells. Mut-GATA1 clones expressed the A gamma* mRNA at the same level as WT clones in both uninduced and induced cells. Thus, GATA-1 binding to this site does not appear to be required for the enhancing function of HS2 in this context. This single-copy rAAV transduction model is useful for evaluating the effects of specific mutations in regulatory elements on the transcription of linked genes.
pubmed:grant
pubmed:commentsCorrections
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
AIM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Sep
pubmed:issn
0006-4971
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
82
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1900-6
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:8400240-Binding Sites, pubmed-meshheading:8400240-DNA-Binding Proteins, pubmed-meshheading:8400240-Dependovirus, pubmed-meshheading:8400240-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:8400240-Enhancer Elements, Genetic, pubmed-meshheading:8400240-Erythroid-Specific DNA-Binding Factors, pubmed-meshheading:8400240-GATA1 Transcription Factor, pubmed-meshheading:8400240-Gene Expression, pubmed-meshheading:8400240-Gene Rearrangement, pubmed-meshheading:8400240-Genetic Vectors, pubmed-meshheading:8400240-Globins, pubmed-meshheading:8400240-Humans, pubmed-meshheading:8400240-Leukemia, Erythroblastic, Acute, pubmed-meshheading:8400240-Mutagenesis, Site-Directed, pubmed-meshheading:8400240-NF-E2 Transcription Factor, pubmed-meshheading:8400240-NF-E2 Transcription Factor, p45 Subunit, pubmed-meshheading:8400240-Polymerase Chain Reaction, pubmed-meshheading:8400240-RNA, Messenger, pubmed-meshheading:8400240-Recombination, Genetic, pubmed-meshheading:8400240-Transcription Factors, pubmed-meshheading:8400240-Tumor Cells, Cultured, pubmed-meshheading:8400240-Zinc Fingers
pubmed:year
1993
pubmed:articleTitle
Single-copy transduction and expression of human gamma-globin in K562 erythroleukemia cells using recombinant adeno-associated virus vectors: the effect of mutations in NF-E2 and GATA-1 binding motifs within the hypersensitivity site 2 enhancer.
pubmed:affiliation
Clinical Hematology Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.