Purification of an Arabidopsis chloroplast extract with in vitro RNA processing activity on psbA and petD 3 '-untranslated regions
Por:
Coloapa-Soto, D, Vargas-Suarez, M, Loza-Tavera, H
Publicada:
1 mar 2012
Resumen:
The mature 3'-end of many chloroplast mRNAs is generated by the processing of the 3'-untranslated region (3'-UTR), which is a mechanism that involves the removal of a segment located downstream an inverted repeat sequence that forms a stem-loop structure. Nuclear-encoded chloroplast RNA binding proteins associate with the stem-loop to process the 3'-UTR or to influence mRNA stability. A spinach chloroplast processing extract (CPE) has been previously generated and used to in vitro dissect the biochemical mechanism underlying 3'-UTR processing. Being Arabidopsis thaliana an important genetic model, the development of a CPE allowing to correlate 3'-UTR processing activity with genes encoding proteins involved in this process, would be of great relevance. Here, we developed a purification protocol that generated an Arabidopsis CPE able to correctly process a psbA 3'-UTR precursor. By UV crosslinking, we characterized the protein patterns generated by the interaction of RNA binding protein
Filiaciones:
Coloapa-Soto, D:
Univ Nacl Autonoma Mexico, Dept Bioquim, Fac Quim, Mexico City 04510, DF, Mexico
Vargas-Suarez, M:
Univ Nacl Autonoma Mexico, Dept Bioquim, Fac Quim, Mexico City 04510, DF, Mexico
Loza-Tavera, H:
Univ Nacl Autonoma Mexico, Dept Bioquim, Fac Quim, Mexico City 04510, DF, Mexico
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