Abstract:
:The cis-requirements for the first editing site in the atp9 mRNA from pea mitochondria were investigated in an in vitro RNA editing system. Template RNAs deleted 5' of -20 are edited correctly, but with decreased efficiency. Deletions between -20 and the edited nucleotide abolish editing activity. Substitution of the sequences 3' of the editing site has little effect, which suggests that the major determinants reside upstream. Stepwise mutated RNA sequences were used as templates or competitors that divide the cis-elements into several distinct regions. In the template RNAs, mutation of the sequence between -40 and -35 reduces the editing activity, while the region from -15 to -5 is essential for the editing reaction. In competition experiments the upstream region can be titrated, while the essential sequence near the editing site is largely resistant to excess competitor. This observation suggests that either one trans-factor attaches to these separate cis-regions with different affinities or two distinct trans-factors bind to these sequences, and one of which is present in limited amounts, whereas the other one is more abundant in the lysate.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Takenaka M,Neuwirt J,Brennicke Adoi
10.1093/nar/gkh763keywords:
subject
Has Abstractpub_date
2004-08-04 00:00:00pages
4137-44issue
14eissn
0305-1048issn
1362-4962pii
32/14/4137journal_volume
32pub_type
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