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Nucleic Acids Research, 1985, Vol. 13, No. 17 6059-6074
© 1985

Articles

Characterization of an unique RNA initiated immediately upstream from human {alpha}1 globin gene in vivo and in vitro: polymerase II-dependence, tissue specificity, and subcellular location

John Hess*, Carlos Perez-Stable*, Al Deisseroth+ and Che-Kun James Shen*,§

*Department of Genetics, University of California, Davis CA 95616 +Department of Medicine, Division of Hemalology and Oncology, University of California San Francisco, CA 94143, USA

§To whom correspondence should be addressed

Received June 17, 1985. Revised August 5, 1985. Accepted August 5, 1985.

We have identified an abundant transcript initiated upstream from the canonical cap site of human {alpha}1 globin gene in bone marrow cells and in COS-7 cells transfected with an {alpha}1 globin gene-containing plasmid. Similar to the major {alpha}1 globin transcript, this upstream RNA is present almost exclusively in the cytoplasm of the transfected COS-7 cells. It is also synthesized efficiently in vitro by RNA polymerase II in the nuclear extracts prepared from a Hela cell line and an erythroleukemia cell line, K562. RNAs isolated from these cell lines, however, do not contain this upstream transcript. The putative 5' end of the {alpha}1 globin upstream RNA is mapped by primer extension to base –45, which is located in between the CCAAT and TATA boxes. The synthesis of this RNA in vitro and in vivo, and the close proximity of its 5' end to the promoter of the {alpha}1 globin gene suggest a common mechanism regulating the transcriptional initiation of both the upstream and the major {alpha}1 globin RNAs.


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