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A fetal globin gene mutation in /sup A/. gamma. nondeletion hereditary persistence of fetal hemoglobin increases promoter strength in a nonerythroid cell

Journal Article · · Mol. Cell. Biol.; (United States)
DOI:https://doi.org/10.1128/MCB.8.2.713· OSTI ID:5115108

Single base substitutions have been identified in the promoter regions of /sup A/..gamma..-globin genes from individuals with certain types of nondeletion /sup A/..gamma.. hereditary persistence of fetal hemoglobin (HPFH). The presence of these mutations is closely associated with the /sup A/..gamma.. HPFH phenotype, but proof that they are the nondeletion HPFH determinants is lacking. To test directly whether these base substitutions can result in an increase in /sup A/..gamma..-globin gene transcription, we studied cosmid clones containing the /sup G/..gamma..-through ..beta..-globin gene regions from individuals with Greek-type (G-to-A base substitution at -117) and Chinese-type (C-to-T base substitution at -196) /sup A/..gamma.. HPFH in a transient expression assay. When tested as part of a cosmid clone, the Greek HPFH /sup A/..gamma..-globin gene consistently produced about 1.4 times as much RNA as the wild-type /sup A/..gamma..-globin gene when standardized against RNA transcribed from the /sup G/..gamma.. genes in cis. The relative strengths of the normal and HPFH /sup A/..gamma..-globin gene promoters were also compared in transient expression assays with plasmids containing the /sup A/..gamma..-globin genes. Pseudo-wild-type /sup A/..gamma..-globin genes containing a short, transcriptionally neutral deletion were used so that two /sup A/..gamma..-globin genes that differed in their promoter sequences could be compared in the same transfection. The plasmid transient expression results indicated a 1.3- to 1.4-fold increase in steady-state RNA levels from the Greek-type /sup A/..gamma.. HPFH promoter compared with the wild-type /sup A/..gamma.. promoter, while no difference was documented between the Chinese-type /sup A/..gamma.. HPFH promoter and the wild-type /sup A/..gamma.. promoter.

Research Organization:
Dow Chemical, U.S.A., Midland, MI 48674 (US)
OSTI ID:
5115108
Journal Information:
Mol. Cell. Biol.; (United States), Journal Name: Mol. Cell. Biol.; (United States) Vol. 8:2; ISSN MCEBD
Country of Publication:
United States
Language:
English