INVESTIGADORES
PALATNIK Javier Fernando
artículos
Título:
Identification of MicroRNA Processing Determinants by Random Mutagenesis of Arabidopsis MIR172a Precursor
Autor/es:
MATEOS, JL; BOLOGNA, NG; CHOROSTECKI, U; PALATNIK JF
Revista:
CURRENT BIOLOGY
Editorial:
CELL PRESS
Referencias:
Año: 2010 p. 49 - 54
ISSN:
0960-9822
Resumen:
MicroRNAs (miRNAs) are widespread posttranscriptional
regulators of gene expression. They are processed from
longer primary transcripts that contain foldback structures
(reviewed in [1, 2]). In animals, a complex formed by Drosha
and DGCR8/Pasha recognizes the transition between the
single-stranded RNA sequences and the stem loop to
produce the first cleavage step in miRNA biogenesis [3].
Whereas animal precursors are of uniform size and shape,
their plant counterparts comprise a collection of variable
stem loops, and little is known about the structural clues
recognized during their processing. Here, we designed an
unbiased approach based on the random mutagenesis of
the MIR172a precursor to study miRNA processing in plants.
Randomly mutated precursors were overexpressed in[1, 2]). In animals, a complex formed by Drosha
and DGCR8/Pasha recognizes the transition between the
single-stranded RNA sequences and the stem loop to
produce the first cleavage step in miRNA biogenesis [3].
Whereas animal precursors are of uniform size and shape,
their plant counterparts comprise a collection of variable
stem loops, and little is known about the structural clues
recognized during their processing. Here, we designed an
unbiased approach based on the random mutagenesis of
the MIR172a precursor to study miRNA processing in plants.
Randomly mutated precursors were overexpressed in[3].
Whereas animal precursors are of uniform size and shape,
their plant counterparts comprise a collection of variable
stem loops, and little is known about the structural clues
recognized during their processing. Here, we designed an
unbiased approach based on the random mutagenesis of
the MIR172a precursor to study miRNA processing in plants.
Randomly mutated precursors were overexpressed inMIR172a precursor to study miRNA processing in plants.
Randomly mutated precursors were overexpressed in
Arabidopsis, and their activity was determined in vivo. We
gathered sequence data from these transgenes and used it
to build a MIR172a precursor map highlighting relevant
and neutral positions for its processing. A 15 nucleotide
stem segment below the miRNA/miRNA* duplex was essential
for MIR172a processing. In contrast, mutations in the
terminal-loop region were mostly neutral, yet a loop was
required for miR172 biogenesis. The results could be
extended to other precursors, suggesting the existence of
common features in at least part of the plant precursors., and their activity was determined in vivo. We
gathered sequence data from these transgenes and used it
to build a MIR172a precursor map highlighting relevant
and neutral positions for its processing. A 15 nucleotide
stem segment below the miRNA/miRNA* duplex was essential
for MIR172a processing. In contrast, mutations in the
terminal-loop region were mostly neutral, yet a loop was
required for miR172 biogenesis. The results could be
extended to other precursors, suggesting the existence of
common features in at least part of the plant precursors.MIR172a precursor map highlighting relevant
and neutral positions for its processing. A 15 nucleotide
stem segment below the miRNA/miRNA* duplex was essential
for MIR172a processing. In contrast, mutations in the
terminal-loop region were mostly neutral, yet a loop was
required for miR172 biogenesis. The results could be
extended to other precursors, suggesting the existence of
common features in at least part of the plant precursors.MIR172a processing. In contrast, mutations in the
terminal-loop region were mostly neutral, yet a loop was
required for miR172 biogenesis. The results could be
extended to other precursors, suggesting the existence of
common features in at least part of the plant precursors.