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Isolation of a U‐insertion/deletion editing complex from Leishmania tarentolae mitochondria
Author(s) -
Aphasizhev Ruslan,
Aphasizheva Inna,
Nelson Robert E.,
Gao Guanghan,
Simpson Agda M.,
Kang Xuedong,
Falick Arnold M.,
Sbicego Sandro,
Simpson Larry
Publication year - 2003
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/cdg083
Subject(s) - biology , rna editing , rna , trypanosoma brucei , exonuclease , guide rna , microbiology and biotechnology , tandem affinity purification , rnase h , multiprotein complex , rna ligase , rnase p , kinetoplast , dna , biochemistry , genetics , affinity chromatography , enzyme , genome , gene , polymerase , cas9
A multiprotein, high molecular weight complex active in both U‐insertion and U‐deletion as judged by a pre‐cleaved RNA editing assay was isolated from mitochondrial extracts of Leishmania tarentolae by the tandem affinity purification (TAP) procedure, using three different TAP‐tagged proteins of the complex. This editing‐ or E‐complex consists of at least three protein‐containing components interacting via RNA: the RNA ligase‐containing L‐complex, a 3′ TUTase (terminal uridylyltransferase) and two RNA‐binding proteins, Ltp26 and Ltp28. Thirteen approximately stoichiometric components were identified by mass spectrometric analysis of the core L‐complex: two RNA ligases; homologs of the four Trypanosoma brucei editing proteins; and seven novel polypeptides, among which were two with RNase III, one with an AP endo/exonuclease and one with nucleotidyltransferase motifs. Three proteins have no similarities beyond kinetoplastids.