期刊
NUCLEIC ACIDS RESEARCH
卷 39, 期 18, 页码 8065-8077出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkr478
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资金
- Plan Nacional I+D+I del Ministerio de Ciencia e Innovacion (MICINN)-Spain [BFU2007/65095/BMC, BFU2007-64999/BMC, BFU2010-16470]
- Instituto de Salud Carlos III (ISCIII)-Redes Tematicas de Investigacion Cooperativa en Salud (RETIC)-Spain [RD06/0021/0014, RD06/0021/0008]
- Fondo Europeo de Desarrollo Regional (FEDER)
L1Tc is a non-LTR LINE element from Trypanosoma cruzi that encodes its transposition machinery and bears an internal promoter. Herewith, we report the identification of an in vitro active hepatitis delta virus-like ribozyme located in the first 77 nt at the 5'-end of the L1Tc mRNA (L1TcRz). The data presented show that L1TcRz has a co-transcriptional function. Using gel-purified uncleaved RNA transcripts, the data presented indicate that the kinetics of the self-cleaving, in a magnesium-dependent reaction, fits to a two-phase decay curve. The cleavage point identified by primer extension takes place at +1 position of the element. The hydroxyl nature of the 5'-end of the 3'-fragment generated by the cleavage activity of L1TcRz was confirmed. Since we have previously described that the 77-nt long fragment located at the 5'-end of L1Tc has promoter activity, the existence of a ribozyme in L1Tc makes this element to be the first described non-LTR retroelement that has an internal promoter-ribozyme dual function. The L1Tc nucleotides located downstream of the ribozyme catalytic motif appear to inhibit its activity. This inhibition may be influenced by the existence of a specific L1Tc RNA conformation that is recognized by RNase P.
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