INVESTIGADORES
BLANCO Flavio Antonio
congresos y reuniones científicas
Título:
Selective recruitment of mRNAs and miRNAs to the translational machinery during Medicago truncatula root nodule organogenesis
Autor/es:
REYNOSO, MAURICIO; BLANCO, FLAVIO; BAILEY-SERRES; CRESPI, MARTÍN; ZANETTI, MARÍA EUGENIA
Lugar:
cold Spring Harbor
Reunión:
Congreso; Cold Spring Harbor 77th Symposium: The Biology of Plants; 2012
Institución organizadora:
Cold Spring Harbor
Resumen:
development or in response to environmental stimuli (1). The symbiotic interaction
between legume and rhizobia involves two highly coordinated processes, the nodule
organogenesis and the bacterial infection. The interaction is initiated by the perception
of the bacteria, which suppresses plant defence responses, infects the roots tissue and
colonizes the developing nodule primordium (2). Several studies have characterized the
steady-state levels of mRNAs at different stages of nitrogen fixing symbiosis. However,
abundance of an mRNA does not necessarily reflect its translation status. In addition,
microRNAs (miRNAs) have emerged as major regulators of mRNA translation or
stability. Hence, we characterized changes in the association of mRNAs and miRNAs to
polyrribosome (polysomes) in roots of the model legume Medicago truncatula during
the symbiotic interaction with Sinorhizobium meliloti. Purification of polysomes was
achieved by immunoprecipitation (IP) using root tissue expressing a FLAG-tagged
ribosomal protein (RP) L18 (3, 4). Quantitative comparison between S. meliloti
achieved by immunoprecipitation (IP) using root tissue expressing a FLAG-tagged
ribosomal protein (RP) L18 (3, 4). Quantitative comparison between S. meliloti
the symbiotic interaction with Sinorhizobium meliloti. Purification of polysomes was
achieved by immunoprecipitation (IP) using root tissue expressing a FLAG-tagged
ribosomal protein (RP) L18 (3, 4). Quantitative comparison between S. meliloti
achieved by immunoprecipitation (IP) using root tissue expressing a FLAG-tagged
ribosomal protein (RP) L18 (3, 4). Quantitative comparison between S. meliloti
Medicago truncatula during
the symbiotic interaction with Sinorhizobium meliloti. Purification of polysomes was
achieved by immunoprecipitation (IP) using root tissue expressing a FLAG-tagged
ribosomal protein (RP) L18 (3, 4). Quantitative comparison between S. meliloti
achieved by immunoprecipitation (IP) using root tissue expressing a FLAG-tagged
ribosomal protein (RP) L18 (3, 4). Quantitative comparison between S. meliloti
Sinorhizobium meliloti. Purification of polysomes was
achieved by immunoprecipitation (IP) using root tissue expressing a FLAG-tagged
ribosomal protein (RP) L18 (3, 4). Quantitative comparison between S. melilotiS. meliloti
inoculated and non inoculated roots revealed differential translational status for genes of
the nodulation signalling pathway. Three translational categories were defined: 1)
transcripts up-regulated at translational level, which includes NFP and CRE1 receptors,
the GRAS family transcription factors NSP1 and NSP2, and the HAP2-1 and HAP5b
subunits of the CCAAT binding factor; 2) translational down-regulated transcripts,
represented by the cation channel DMI1 and 3) transcripts not regulated at translational
level, including the transcription factors NIN and ERN, the early nodulin ENOD40, the
LYK3 receptor and the calcium calmodulin dependent kinase DMI3. Quantitative
analysis of sRNAs associated with the immunopurified polysome detected several
mature microRNAs (miRNAs), notably miR169d, which targets the 3`UTR of the
HAP2-1 transcript (5). Association of this miRNA with polysomes significantly
decreased upon inoculation with S. meliloti in correlation with increased levels of
HAP2-1 protein. Current experiments are being conducted to explore translational
changes at genome scale combining the IP of polysomes with RNA sequencing
technology.
1) Bailey Serres et al (2009). Trends in Plant Sci 14, 443.
2) Oldroyd et al (2011). Annu Rev Genet 45, 119.
3) Zanetti et al (2005) Plant Phys 138, 624
4) Mustroph et al, (2009) PNAS USA 106, 18843
5) Devers et al, 2011. Plant Phys 156, 1990.