Structural determinants of Arabidopsis thaliana hyponastic leaves 1 function in vivo
Burdisso, Paula (Instituto de Biología Molecular y Celular de Rosario)
Milia, Fernando (Instituto de Biología Molecular y Celular de Rosario)
Schapire, Arnaldo L. (Centre de Recerca en Agrigenòmica)
Bologna, Nicolás (Instituto de Biología Molecular y Celular de Rosario)
Palatnik, Javier F. (Instituto de Biología Molecular y Celular de Rosario)
Rasia, Rodolfo M. (Instituto de Biología Molecular y Celular de Rosario)

Data:	2014
Resum:	MicroRNAs have turned out to be important regulators of gene expression. These molecules originate from longer transcripts that are processed by ribonuclease III (RNAse III) enzymes. Dicer proteins are essential RNAse III enzymes that are involved in the generation of microRNAs (miRNAs) and other small RNAs. The correct function of Dicer relies on the participation of accessory dsRNA binding proteins, the exact function of which is not well-understood so far. In plants, the double stranded RNA binding protein Hyponastic Leaves 1 (HYL1) helps Dicer Like protein (DCL1) to achieve an efficient and precise excision of the miRNAs from their primary precursors. Here we dissected the regions of HYL1 that are essential for its function in Arabidopsis thaliana plant model. We generated mutant forms of the protein that retain their structure but affect its RNA-binding properties. The mutant versions of HYL1 were studied both in vitro and in vivo, and we were able to identify essential aminoacids/residues for its activity. Remarkably, mutation and even ablation of one of the purportedly main RNA binding determinants does not give rise to any major disturbances in the function of the protein. We studied the function of the mutant forms in vivo, establishing a direct correlation between affinity for the pri-miRNA precursors and protein activity.
Ajuts:	Ministerio de Ciencia e Innovación PICT2008/318 Ministerio de Ciencia e Innovación PICT2007/720
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Publicat a:	PloS one, Vol. 9, issue 11 (November 2014) , e113243, ISSN 1932-6203

DOI: 10.1371/journal.pone.0113243
PMID: 25409478

8 p, 1.4 MB

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