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J Biol Chem, Vol. 275, Issue 10, 7261-7272, March 10, 2000

Human Transaldolase-associated Repetitive Elements Are Transcribed by RNA Polymerase III^*

Andras Perl, Emanuela Colombo, Ella Samoilova, Mark C. Butler, and Katalin Banki

From the Departments of Medicine, Microbiology and Immunology, and Pathology, State University of New York Health Science Center, College of Medicine, Syracuse, New York 13210

Repetitive elements flanked by exons 2 and 3 of the human transaldolase gene, thus termed transaldolase-associated repetitive elements, TARE, were identified in human DNA. Nonpolyadenylated TARE transcripts were detected by Northern blot analysis and cloned by reverse transcriptase-mediated polymerase chain reaction from human T lymphocytes. A dominant 1085-nucleotide long transcript, TARE-6, contained two adjacent Alu elements, a right monomer and a complete dimer, oriented opposite to the direction of transcription of the transaldolase gene. Reverse transcriptase-polymerase chain reaction and in vitro transcription analyses showed that transcription of TARE-6 proceeded in the orientation of the RNA pol III promoter of the Alu dimer and opposite to the orientation of the TAL-H gene. TAREs lacking RNA polymerase III promoter showed no transcriptional activity. In vitro transcription of TARE-6 was resistant to 1 µg/ml -amanitin but sensitive to 100 µg/ml -amanitin and tagetitoxin, suggesting involvement of RNA polymerase III. TAREs in both the transaldolase and HSAG-1 genomic loci were surrounded by TA target site duplications. Homologies between transaldolase and HSAG-1 break off internally at splice donor and acceptor sites. The results suggest RNA polymerase III-mediated transcription of TARE may be a source of repetitive elements, contributing to distinct genes and thus shaping the human genome.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EMBL Data Bank with accession number(s) L19437 (TAL-H cDNA), AF058913 (TAL-H genomic DNA locus), L27346 (TARE-6), and X03822 (HSAG-1).

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