INVESTIGADORES
ZANETTI Maria Eugenia
congresos y reuniones científicas
Título:
COMBINING IMMUNOPURIFICATION OF RIBONUCLEOPROTEIN COMPLEXES AND RNA-SEQ TO CHARACTERIZE POSTRANSCRIPCIONAL GENE REGULATION IN THE SYMBIOTIC INTERACTION Medicago truncatula- Sinorhizobium meliloti
Autor/es:
MARCOS LANCIA, MAURICIO REYNOSO, FLAVIO BLANCO Y MARÍA EUGENIA ZANETTI
Lugar:
Chascomús
Reunión:
Congreso; XV Jornadas Anuales de la Sociedad Argentina de Biología; 2013
Institución organizadora:
Sociedad Argentina de Biología
Resumen:
Several
studies have characterized the steady-state levels of transcripts at different
stages of root legume symbiosis. However, this appoach does not
distinguish transcripts that are targeted for
degradation, sequestered in ribonucleoprotein complexes or undergoing
translation. The distribution of mRNAs on these cytoplasmic complexes
plays a crucial role in the regulation of gene expression during development or
in response to environmental stimuli. Here, we characterized changes in the
association of transcripts to polysomes in roots of the model legume Medicago truncatula during the symbiotic
interaction with Sinorhizobium meliloti.
Polysomes were isolated by immunopurification (IP) from root tissue expressing
a FLAG-tagged ribosomal protein L18. This methodology was combined with
Illumina RNA sequencing technology (RNA-seq) allowing a quantitative analysis
at genome scale of inoculated and control roots. Approximately 3500 transcripts
were classified as up-regulated at translational level, whereas 5770
transcripts were found as down-regulated. A comparison of fold change levels in
response to rhizobia of 12 transcripts encoding proteins involved in the
nodulation signalling pathway obtained previously by qRT-PCR showed a good
correlation with data generated by RNA-seq. Current experiments are being
conducted to immunopurify other cytoplasmic complexes involved in storage or
degradation of mRNAs, such as storage
granules or P Bodies. This
experimental approach constitutes a valuable tool to shed light on the
mechanisms and significance of post-transcriptional regulation in M. truncatula roots during its
association with S. meliloti.