PKR activation by a non-coding RNA expressed in lymphocytes of mice bearing B16 melanoma.
In recent years, non-coding RNAs (ncRNAs) have become an exciting area of research. It has been demonstrated that ncRNAs play an important role in the regulation of gene expression in eukaryotic cells. However, little is known about ncRNAs in lymphocytes. In this study, we investigated the presence of ncRNAs in lymphocytes of C57BL/6 mice bearing B16 melanoma by using the differential display reverse transcription-PCR (DD-RT-PCR). PKR is a serine/threonine kinase containing two RNA-binding domains within the N-terminal region. We took advantage of the ability of RNAs to bind PKR in order to identify ncRNAs of lymphocytes activated during tumor development. Thus, RNAs that co-immunoprecipitated with PKR were reversed transcribed, re-amplified, cloned, sequenced and the secondary structure was determined. The ability of transcripts obtained by in vitro transcription to activate PKR was also examined. We detected a highly structured transcript of 220 nt with no open reading frame (ORF) which is able to activate PKR, and it is only expressed in lymphocytes of C57BL/6 mice bearing B16 melanoma. Therefore, the 220 nt transcript may be included in the class of ncRNAs that act by modifying protein activity and our data suggest that regulation of gene expression in activated lymphocytes by this ncRNA could be mediated by PKR through the activation of the transcription factor NF-kappaB.
Murad
Joana M
JM
Department of Biochemistry and Immunology, School of Medicine University of São Paulo, 14049-900 Ribeirão Preto, SP, Brazil.
de Souza
Liliana R
LR
De Lucca
Fernando L
FL
D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D004789:Enzyme Activation; D008214:Lymphocytes / Q000378:metabolism*; D008546:Melanoma, Experimental / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000378:metabolism*; D022661:RNA, Untranslated / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment; D014407:Tumor Cells, Cultured; D019892:eIF-2 Kinase / Q000378:metabolism*
D022661:RNA, Untranslated; D019892:eIF-2 Kinase