Mutations in two regions upstream of the A gamma globin gene canonical promoter affect gene expression.
Nucleic Acids Res
; 17(11): 4339-52, 1989 Jun 12.
Article
in En
| MEDLINE
| ID: mdl-2472607
ABSTRACT
Two regions upstream of the human fetal (A gamma) globin gene, which interact with protein factors from K562 and HeLa nuclear extracts, have functional significance in gene expression. One binding site (site I) is at a position -290 to -267 bp upstream of the transcription initiation site, the other (site II) is at -182 to -168 bp. Site II includes the octamer sequence (ATGCAAAT) found in an immunoglobulin enhancer and the histone H2b gene promoter. A point mutation (T----C) at -175, within the octamer sequence, is characteristic of a naturally occurring HPFH (hereditary persistence of fetal hemoglobin), and decreases factor binding to an oligonucleotide containing the octamer motif. Expression assays using a A gamma globin promoter-CAT (chloramphenicol acetyl transferase) fusion gene show that the point mutation at -175 increases expression in erythroid, but not non-erythroid cells when compared to a wild-type construct. This correlates with the actual effect of the HPFH mutation in humans. This higher expression may result from a mechanism more complex than reduced binding of a negative regulator. A site I clustered-base substitution gives gamma-CAT activity well below wild-type, suggesting that this factor is a positive regulator.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Fetal Hemoglobin
/
Globins
/
Gene Expression Regulation
/
Promoter Regions, Genetic
/
Mutation
Limits:
Humans
Language:
En
Journal:
Nucleic Acids Res
Year:
1989
Document type:
Article