PCR-based detection of Pol III-transcribed transposons and its application to the rodent model of ultraviolet response.
Cellular levels of RNAs containing transposable elements increase in response to various stresses. Polymerase III (Pol III)-dependent transcripts of transposons are different from transposon-containing RNAs generated by read-through Pol II-dependent transcription. Until now, Pol III transcripts were detected by primer extension followed by time-consuming gel electrophoresis. In this paper, we describe a more sensitive PCR-based method for the selective detection of Pol III-transcribed RNAs. The method is based on the difference in sequences at the 5' ends of the Pol II- and Pol III-dependent transcripts. We employed this method to quantify Pol III transcripts of transposon B1 in rodent cells and revealed that their levels are affected by UV irradiation. We therefore conclude that the abundance of the Pol III-transcribed fraction of cellular RNA may serve as marker of stress response and can be conveniently quantified by the method described.
Myakishev
Max
M
Department of Dermatology, University of Rochester Medical Center, Rochester, NY 14642, USA. Max_Myakishev@urmc.rochester.edu
Polesskaya
Oksana
O
Kulichkova
Valentina
V
Baranova
Ancha
A
Gause
Larissa
L
Konstantinova
Irina
I
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured / Q000378:metabolism / Q000528:radiation effects; D004258:DNA Polymerase III / Q000378:metabolism*; D004251:DNA Transposable Elements / Q000235:genetics* / Q000528:radiation effects; D020869:Gene Expression Profiling / Q000379:methods*; D022781:Hepatocytes / Q000737:chemistry* / Q000201:enzymology; D015603:Keratinocytes / Q000201:enzymology / Q000378:metabolism / Q000528:radiation effects; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA / Q000096:biosynthesis* / Q000235:genetics; D051381:Rats; D020086:Short Interspersed Nucleotide Elements / Q000235:genetics; D013312:Stress, Physiological / Q000235:genetics*; D014466:Ultraviolet Rays / Q000009:adverse effects*
D004251:DNA Transposable Elements; D012313:RNA; D004258:DNA Polymerase III