In vivo RNA-RNA duplexes from human alpha3 and alpha5 nicotinic receptor subunit mRNAs.
Natural antisense transcripts, because of their potential to form double-stranded RNA (dsRNA) molecules, recently emerged as a mechanism acting on eukaryotic gene regulation at multiple levels. CHRNA3 and CHRNA5, coding for alpha3 and alpha5 subunits of the neuronal nicotinic acetylcholine receptor, have been reported to overlap at their 3'ends in human and bovine genomes. In the present paper, four CHRNA3 and three CHRNA5 human transcripts were characterised, leading to the identification of different antisense complementary regions. Since the two genes are coexpressed in some neuronal and non-neuronal tissues, we ventured on the in vivo identification of RNA-RNA duplexes in both humans and cattle. Using an RNase protection-based approach, CHRNA3/CHRNA5 duplexes were detected in human neuroblastoma SY5Y cells, but not in bovine cerebellum. A semi-quantitative analysis of overlapping transcript levels was performed by real-time RT-PCR. Possible consequences of sense-antisense interaction are discussed.
Soldà
Giulia
G
Department of Biology and Genetics for Medical Sciences, University of Milan, via Viotti 3/5, 20133 Milan, Italy.
Boi
Silvia
S
Duga
Stefano
S
Fornasari
Diego
D
Benfante
Roberta
R
Malcovati
Massimo
M
Tenchini
Maria Luisa
ML
D000818:Animals; D002417:Cattle; D006801:Humans; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D016372:RNA, Antisense; D012333:RNA, Messenger / Q000378:metabolism; D011978:Receptors, Nicotinic / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D009419:Nerve Tissue Proteins; D016372:RNA, Antisense; D012333:RNA, Messenger; D011978:Receptors, Nicotinic; C447158:CHRNA5 protein, human