It has been demonstrated that K562 cells express epsilon- and gamma-globin genes but do not express beta-globin genes. The regulation of gene expression occurs at the level of transcription. We have set up a cell-free in vitro transcription system from K562 cells to determine the requirements for globin mRNA synthesis. As an initial attempt, we have prepared extracts from nuclei of both hemin-induced and uninduced K562 cells. The nuclear extracts could direct accurate transcription initiation in vitro from the epsilon-globin gene promoter without supplement with whole cell extracts. The results up to the present point to the existence of a globin gene expression regulatory factor(s) in K562 nuclear extracts. The in vitro transcription system will be used as an assay system for the isolation and characterization of such factors.

Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
1986
Total Cost
Indirect Cost
Name
U.S. National Inst Diabetes/Digst/Kidney
Department
Type
DUNS #
City
State
Country
United States
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