TY - JOUR
T1 - Minimal pre-mRNA substrates with natural and converted sites for full-round U insertion and U deletion RNA editing in trypanosomes
AU - Cifuentes-Rojas, Catherine
AU - Halbig, Kari
AU - Sacharidou, Anastasia
AU - De Nova-Ocampo, Monica
AU - Cruz-Reyes, Jorge
N1 - Funding Information:
We thank Drs. Linda Guarino, Andy C. LiWang and C.T. Ranjith-Kumar for comments on the manuscript and helpful discussions. This work was supported by a grant from the NIH (GM067130) to JC-R. Funding to pay the Open Access publication charges for this article was provided by the same grant above.
PY - 2005
Y1 - 2005
N2 - Trypanosome RNA editing by uridylate insertion or deletion cycles is a mitochondrial mRNA maturation process catalyzed by multisubunit complexes. A full-round of editing entails three consecutive steps directed by partially complementary guide RNAs: pre-mRNA cleavage, U addition or removal, and ligation. The structural and functional composition of editing complexes is intensively studied, but their molecular interactions in and around editing sites are not completely understood. In this study, we performed a systematic analysis of distal RNA requirements for full-round insertion and deletion by purified editosomes. We define minimal substrates for efficient editing of A6 and CYb model transcripts, and established a new substrate, RPS12. Important differences were observed in the composition of substrates for insertion and deletion. Furthermore, we also showed for the first time that natural sites can be artificially converted in both directions: from deletion to insertion or from insertion to deletion. Our site conversions enabled a direct comparison of the two editing kinds at common sites during substrate minimization and demonstrate that all basic determinants directing the editosome to carry out full-round insertion or deletion reside within each editing site. Surprisingly, we were able to engineer a deletion site into CYb, which exclusively undergoes insertion in nature.
AB - Trypanosome RNA editing by uridylate insertion or deletion cycles is a mitochondrial mRNA maturation process catalyzed by multisubunit complexes. A full-round of editing entails three consecutive steps directed by partially complementary guide RNAs: pre-mRNA cleavage, U addition or removal, and ligation. The structural and functional composition of editing complexes is intensively studied, but their molecular interactions in and around editing sites are not completely understood. In this study, we performed a systematic analysis of distal RNA requirements for full-round insertion and deletion by purified editosomes. We define minimal substrates for efficient editing of A6 and CYb model transcripts, and established a new substrate, RPS12. Important differences were observed in the composition of substrates for insertion and deletion. Furthermore, we also showed for the first time that natural sites can be artificially converted in both directions: from deletion to insertion or from insertion to deletion. Our site conversions enabled a direct comparison of the two editing kinds at common sites during substrate minimization and demonstrate that all basic determinants directing the editosome to carry out full-round insertion or deletion reside within each editing site. Surprisingly, we were able to engineer a deletion site into CYb, which exclusively undergoes insertion in nature.
UR - http://www.scopus.com/inward/record.url?scp=29144533269&partnerID=8YFLogxK
U2 - 10.1093/nar/gki943
DO - 10.1093/nar/gki943
M3 - Artículo
C2 - 16306234
SN - 0305-1048
VL - 33
SP - 6610
EP - 6620
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 20
ER -