A putative RNA editing from U to C in a mouse mitochondrial transcript

Jaime Villegas, Ilse Müller, Jacob Arredondo, Rodrigo Pinto, Luis O. Burzio

Resultado de la investigación: Article

18 Citas (Scopus)

Resumen

Recently, we isolated and characterized a new mouse mitochondrial RNA molecule containing the mitochondrial 16S RNA plus 121 nt joined to the 5′ end of the RNA. This fragment arises from the L strand of the same gene and we have named this transcript chimeric RNA. At position 121 of the RNA there is a C, which, according to the sequence of the mitochondrial 16S RNA gene, should be a U. We hypothesized that this RNA is synthesized having a U at position 121, which is later substituted to a C by a putative editing reaction. Based on the presence of sites for the restriction endonucleases Rsal and Fnu4HI around position 121, both forms of the RNA were detected in mouse tissues. To confirm the presence of the non-edited and putative edited RNA, a fragment containing the first 154 nt of the RNA was amplified by RT-PCR and cloned. The substitution of U for C was demonstrated by sequencing these clones. In vitro transcription experiments demonstrated that the substitution of U for C is not due to artifact of amplification or cloning. Moreover, in mitochondria from testis only the non-edited form was found. This, together with other experimental evidence, demonstrated that the base substitution was not due to polymorphism of the mitochondrial 16S RNA gene. This is the first demonstration of a substitution reaction from U to C in a mammalian mitochondrial transcript.

Idioma originalEnglish
Páginas (desde-hasta)1895-1901
Número de páginas7
PublicaciónNucleic Acids Research
Volumen30
N.º9
EstadoPublished - 1 may 2002

Huella dactilar

RNA Editing
RNA
Genes
DNA Restriction Enzymes
Artifacts
Testis
Organism Cloning
Mitochondria
Clone Cells
Polymerase Chain Reaction
mitochondrial RNA

ASJC Scopus subject areas

  • Genetics

Citar esto

Villegas, J., Müller, I., Arredondo, J., Pinto, R., & Burzio, L. O. (2002). A putative RNA editing from U to C in a mouse mitochondrial transcript. Nucleic Acids Research, 30(9), 1895-1901.
Villegas, Jaime ; Müller, Ilse ; Arredondo, Jacob ; Pinto, Rodrigo ; Burzio, Luis O. / A putative RNA editing from U to C in a mouse mitochondrial transcript. En: Nucleic Acids Research. 2002 ; Vol. 30, N.º 9. pp. 1895-1901.
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Villegas, J, Müller, I, Arredondo, J, Pinto, R & Burzio, LO 2002, 'A putative RNA editing from U to C in a mouse mitochondrial transcript', Nucleic Acids Research, vol. 30, n.º 9, pp. 1895-1901.

A putative RNA editing from U to C in a mouse mitochondrial transcript. / Villegas, Jaime; Müller, Ilse; Arredondo, Jacob; Pinto, Rodrigo; Burzio, Luis O.

En: Nucleic Acids Research, Vol. 30, N.º 9, 01.05.2002, p. 1895-1901.

Resultado de la investigación: Article

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AU - Müller, Ilse

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AU - Pinto, Rodrigo

AU - Burzio, Luis O.

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N2 - Recently, we isolated and characterized a new mouse mitochondrial RNA molecule containing the mitochondrial 16S RNA plus 121 nt joined to the 5′ end of the RNA. This fragment arises from the L strand of the same gene and we have named this transcript chimeric RNA. At position 121 of the RNA there is a C, which, according to the sequence of the mitochondrial 16S RNA gene, should be a U. We hypothesized that this RNA is synthesized having a U at position 121, which is later substituted to a C by a putative editing reaction. Based on the presence of sites for the restriction endonucleases Rsal and Fnu4HI around position 121, both forms of the RNA were detected in mouse tissues. To confirm the presence of the non-edited and putative edited RNA, a fragment containing the first 154 nt of the RNA was amplified by RT-PCR and cloned. The substitution of U for C was demonstrated by sequencing these clones. In vitro transcription experiments demonstrated that the substitution of U for C is not due to artifact of amplification or cloning. Moreover, in mitochondria from testis only the non-edited form was found. This, together with other experimental evidence, demonstrated that the base substitution was not due to polymorphism of the mitochondrial 16S RNA gene. This is the first demonstration of a substitution reaction from U to C in a mammalian mitochondrial transcript.

AB - Recently, we isolated and characterized a new mouse mitochondrial RNA molecule containing the mitochondrial 16S RNA plus 121 nt joined to the 5′ end of the RNA. This fragment arises from the L strand of the same gene and we have named this transcript chimeric RNA. At position 121 of the RNA there is a C, which, according to the sequence of the mitochondrial 16S RNA gene, should be a U. We hypothesized that this RNA is synthesized having a U at position 121, which is later substituted to a C by a putative editing reaction. Based on the presence of sites for the restriction endonucleases Rsal and Fnu4HI around position 121, both forms of the RNA were detected in mouse tissues. To confirm the presence of the non-edited and putative edited RNA, a fragment containing the first 154 nt of the RNA was amplified by RT-PCR and cloned. The substitution of U for C was demonstrated by sequencing these clones. In vitro transcription experiments demonstrated that the substitution of U for C is not due to artifact of amplification or cloning. Moreover, in mitochondria from testis only the non-edited form was found. This, together with other experimental evidence, demonstrated that the base substitution was not due to polymorphism of the mitochondrial 16S RNA gene. This is the first demonstration of a substitution reaction from U to C in a mammalian mitochondrial transcript.

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Villegas J, Müller I, Arredondo J, Pinto R, Burzio LO. A putative RNA editing from U to C in a mouse mitochondrial transcript. Nucleic Acids Research. 2002 may 1;30(9):1895-1901.