Inverted Gcg/CGC trinucleotide microsatellites in the 5'-region of Mus IDS mRNA: recurrent induction of aberrant reverse transcripts.
An investigation was initiated to explore previously published results indicating that approximately 80 bp of the 5'-end of the iduronate sulfatase (IDS) cDNA sequence (Accession No L07291) are 100% homologous with the 3'-UTR of isoform I of the sodium hydrogen exchanger (Acc. No. U51112). 5'-RACE carried out on IDS mRNA demonstrated the apparent homology to be a cloning artifact. A sequence comparison of the IDS 5'-RACE product with a mouse BAC clone covering the region, and with various IDS ESTs, suggested that the region is highly susceptible to cloning artifacts, a common one of which is template switching by reverse transcriptase. The nucleotide sequence flanking the translation start site is unusual in containing two inverted repeats composed of the complementary trinucleotide microsatellites, (GCG)9 and (CGC)6. These likely form a highly stable stem of 20-21 nt, through which reverse transcription is compromised. Such a stem could be involved in the regulation of IDS expression by directly affecting translation, message turnover, or serving as a substrate for siRNA production. Though such mRNA features are relatively rare, they may be more abundant but overlooked due to difficulties in their reverse transcription.
Lobo-Menendez
Fe
F
Department of Biological Sciences, University of South Carolina, Columbia, SC 29208, USA.
Bowman
Lewis H
LH
Dewey
Michael J
MJ
D020121:5' Untranslated Regions / Q000737:chemistry* / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D016477:Artifacts; D001483:Base Sequence; D020224:Expressed Sequence Tags; D006801:Humans; D007066:Iduronate Sulfatase / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism; D048348:Reverse Transcription / Q000235:genetics; D013698:Templates, Genetic; D018911:Trinucleotide Repeats / Q000235:genetics*
D020121:5' Untranslated Regions; D012333:RNA, Messenger; D007066:Iduronate Sulfatase