Upstream flanking sequences and transcription of SINEs

• Published in: Journal of molecular biology Vol. 302; no. 1; pp. 17 - 25
• Main Authors: Roy, A M, West, N C, Rao, A, Adhikari, P, Alemán, C, Barnes, A P, Deininger, P L
• Format: Journal Article
• Language: English
• Published: England 08-09-2000
• Subjects: Alu Elements - genetics; Animals; BC1 gene; Cell Line; DNA, Recombinant - genetics; Evolution, Molecular; Gene Expression Profiling; Gene Expression Regulation - genetics; Humans; Mice; Mice, Inbred C57BL; Mice, Transgenic; Regulatory Sequences, Nucleic Acid - genetics; Retroelements - genetics; Ribonucleoproteins - metabolism; RNA - biosynthesis; RNA - genetics; RNA - metabolism; RNA 7SL; RNA Polymerase III - metabolism; Short Interspersed Nucleotide Elements - genetics; snRNA U6; Transcription, Genetic - genetics; Transgenes - genetics; Tumor Cells, Cultured; Up-Regulation - genetics
• ISSN: 0022-2836
• DOI: 10.1006/jmbi.2000.4027

SINEs, short interspersed repeated DNA elements, undergo amplification through retroposition and subsequent integration into a new location in the genome. Each new SINE insertion will be located in a new chromosomal environment, with different flanking sequences. Modulation of transcription by different flanking sequences may play an important role in determining which SINE elements are preferentially active in a genome. We evaluated the ability of upstream flanking sequences to regulate the transcription of three different SINEs (Alu, B2 and ID) by constructing chimeric constructs with known 5' flanking sequences of RNA polymerase III-transcribed genes. Upstream sequences from the 7SL RNA gene, U6 RNA gene, vault RNA gene, and BC1 gene increase transcription of Alu, B2 and BC1 in transient transfections of NIH3T3, HeLa, Neuro2a and C6 glioma cell lines. The 7SL sequence proved most efficient in increasing SINE transcription. The 7SL upstream fused to the BC1 RNA gene (an ID element) was used to create a transgenic mouse line. In contrast to the tissue-specific endogenous BC1 transcription, BC1 transgene transcripts were detected in all tissues tested. However, expression was much higher in those tissues that express the endogenous gene, demonstrating both transcriptional and post-transcriptional regulation. The BC1 RNA was detected in a similar ribonucleoprotein complex in the different tissues.