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Nucleic Acids Research, 1994, Vol. 22, No. 6 1087-1095
© 1994


MOLECULAR BIOLOGY

Alu transcripts: cytoplasmic localisation and regulation by DNA methylation

Wen-Man Liu1, Richard J. Maraia3, Carol M. Rubin1 and Carl W. Schmid1,2,*

1Section of Molecular and Cellular Biology, University of California Davis, CA 95616 2Department of Chemistry, University of California Davis, CA 95616 3National Institute of Child Health and Human Development, NIH Bethesda, MD 20892, USA

*To whom correspondence should be addressed

Received September 30, 1993. Revised February 7, 1994. Accepted February 7, 1994.

Full length Alu transcripts in HeLa cells are detected by primer extension using reverse transcriptase and are also analyzed as cloned cDNA sequences. The 5' end of these transcripts corresponds to the transcriptional start site for RNA polymerase III indicating that these RNAs are transcribed from their internal polymerase III promoters. The Alu transcripts found in cytoplasmic poly A+ RNAs appear to be organized into RNPs as assayed by sucrose gradient sedimentation. Present at about one hundred to one thousand copies per cell, the Alu transcripts are rare as compared to 7SL RNA. In agreement with previous reports that methylation inhibits Pol Ill-directed transcription of Alu in vitro, treatment of HeLa cells with 5-azacytldine results in Alu DNA hypomethylation and an increase in the abundance of the Alu transcript. Sequence analysis shows that many different Alu repeats including members of all subfamilies are transcribed by Pol III In vivo. cDNA sequences of the Pol Ill-directed transcripts exactly match the A box of the Pol III promoter element whereas in other Alu transcripts this element is not faithfully conserved.


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