Cloning and characterization of two guide RNA-binding proteins from mitochondria of Crithidia fasciculata: gBP27, a novel protein, and gBP29, the orthologue of Trypanosoma brucei gBP21

Blom, Daniël; Burg, J. van den; Breek, C. K. D.; Speijer, Dave; Muijsers, Anton O.; Benne, Rob

doi:10.1093/nar/29.14.2950

Cited by 51 publications

(51 citation statements)

References 63 publications

(65 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The L. tarentolae Ltp26 sequence showed strong homology with C. fasciculata gBP27 (Blom et al 2001). The L. tarentolae Ltp26 protein also produced a significant hit with an EST clone (AI881040) in the T. brucei Genome Database.…”

Section: Isolation Of Ltp26 and Ltp28 Proteins And Cloning Of Genesmentioning

confidence: 97%

“…A BLAST search revealed a strong homology of Ltp28 with T. brucei gBP21 (Koller et al 1997) and C. fasciculata gBP29 (Blom et al 2001). The L. tarentolae Ltp26 sequence showed strong homology with C. fasciculata gBP27 (Blom et al 2001).…”

Section: Isolation Of Ltp26 and Ltp28 Proteins And Cloning Of Genesmentioning

confidence: 99%

“…In Crithidia fasciculata, gBP29, an ortholog of gBP21, and a second related RNA-binding protein, gBP27, were cloned. In addition, an ortholog of gPB27 from T. brucei, gBP25, was identified (Leegwater et al 1995;Blom et al 2001). These proteins from C. fasciculata coimmunoprecipitated and were present in the form of variable-sized high-molecular-weight complexes in the mitochondrial extract.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

A 100-kD complex of two RNA-binding proteins from mitochondria of Leishmania tarentolae catalyzes RNA annealing and interacts with several RNA editing components

Aphasizhev¹,

Aphasizheva²,

Nelson³

et al. 2003

RNA

101

121

View full text Add to dashboard Cite

A stable 100-kD complex from mitochondria of Leishmania tarentolae containing two RNA-binding proteins, Ltp26 and Ltp28, was identified by cross-linking to unpaired 4-thiouridine nucleotides in a partially duplex RNA substrate. The genes were cloned and expressed and the complex was reconstituted from recombinant proteins in the absence of RNA or additional factors. The Ltp26 and Ltp28 proteins are homologs of gBP27 and gBP29 from Crithidia fasciculata and gBP25 and gBP21 from Trypanosoma brucei, respectively. The purified Ltp26/Ltp28 complex, the individual recombinant proteins, and the reconstituted complex are each capable of catalyzing the annealing of complementary RNAs, as was previously shown for gBP21 from T. brucei. A high-molecular-weight RNP complex consisting of the Ltp26/Ltp28 complex and several 55-60-kD proteins together with guide RNA could be purified from mitochondrial extract of L. tarentolae transfected with Ltp28-TAP. This complex also interacted in a less stable manner with the RNA ligase-containing L-complex and with the 3 TUTase. The Ltp26/Ltp28 RNP complex is a candidate for catalyzing the annealing of guide RNA and pre-edited mRNA in the initial step of RNA editing.

show abstract

Section: Isolation Of Ltp26 and Ltp28 Proteins And Cloning Of Genesmentioning

confidence: 97%

Section: Isolation Of Ltp26 and Ltp28 Proteins And Cloning Of Genesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

A 100-kD complex of two RNA-binding proteins from mitochondria of Leishmania tarentolae catalyzes RNA annealing and interacts with several RNA editing components

Aphasizhev¹,

Aphasizheva²,

Nelson³

et al. 2003

RNA

101

121

View full text Add to dashboard Cite

show abstract

“…The mitochondrial RNA-binding protein complex contains two small RNA-binding proteins and three associated proteins, and may possibly be involved in the initial annealing of the guide RNA and the mRNA (4)(5)(6)(7)(8)(9). The RNAediting terminal uridylyl transferase 1 (TUTase 1) (RET1) complex contains the RET1 tetramer and one or more unidentified components (10).…”

mentioning

confidence: 99%

Functional complementation of Trypanosoma brucei RNA in vitro editing with recombinant RNA ligase

Gao

Simpson

Kang

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The Ϸ20S RNA ligase-containing complex (L-complex) in trypanosomatid mitochondria interacts by means of RNA linkers with at least two other multiprotein complexes to mediate the editing of mitochondrial cryptogene transcripts. The L-complex contains Ϸ16 proteins, including the two RNA-editing ligases (RELs), REL1 and REL2. Leishmania tarentolae REL1 and REL2 and Trypanosoma brucei REL1 were expressed as enzymatically active tandem affinity purification-tagged proteins in a Baculovirus system. When these proteins were added to mitochondrial lysates from T. brucei procyclic cells that were depleted of the cognate endogenous ligase by RNA interference down-regulation of expression, the added proteins were integrated into the L-complex, and, in the case of REL1, there was a complementation of in vitro-precleaved U-insertion and U-deletion editing activities of the 20S L-complex. Integration of the recombinant proteins did not occur or occurred at a very low level with noncognate ligase-depleted L-complex or with wild-type L-complex. A C-terminal region of the T. brucei recombinant REL1 downstream of the catalytic domain was identified as being involved in integration into the L-complex. The ability to perform functional complementation in vitro provides a powerful tool for molecular dissection of the editing reaction.editing ͉ RNA-editing ligase 1 ͉ RNA-editing ligase 2 ͉ trypanosome S everal macromolecular complexes have been identified that are involved in U-insertion͞deletion RNA editing in trypanosomatid mitochondria (1-3). The mitochondrial RNA-binding protein complex contains two small RNA-binding proteins and three associated proteins, and may possibly be involved in the initial annealing of the guide RNA and the mRNA (4-9). The RNAediting terminal uridylyl transferase 1 (TUTase 1) (RET1) complex contains the RET1 tetramer and one or more unidentified components (10). RET1 has been shown to be involved in the addition of nonencoded Us to the 3Ј ends of guide RNAs (11). The RNA ligase-containing L-complex was initially detected as an ␣-32 P-ATP-labeled particle sedimenting Ϸ20S in glycerol gradients and migrating as a single band with an approximate size of 1,200-2,000 kDa in native gels (12,13). This complex has been labeled the editosome but it seems appropriate to reserve this nomenclature for the entire RNA͞protein supercomplex once it is more defined. The L-complex has been isolated from Trypanosoma brucei and Leishmania tarentolae mitochondria by column chromatography (13-16) and by tandem affinity purification (TAP) chromatography (17). At least 16 protein components have been identified, including the two RNA-editing ligases (RELs) (REL1 and REL2), a second 3Ј TUTase (RET2), three to four RNase III-motif proteins, three related zinc finger-motif proteins, two AP-endo-exonucleasephosphatase-motif proteins, two RNA binding-motif proteins, and several proteins lacking identifiable motifs.The two ligases are localized in subcomplexes that have been partially defined by coimmunoprecipitation experiments and...

show abstract

“…Some other proteins may play roles in editing while not being a stable part of the editosome. These may include a 108 kDa protein with TUTase activity that is essential for editing [12], and RNA binding proteins gBP21 [13] and related protein gBP25 ( [14] and J. Lukes personal communication), and other RNA binding proteins RBP16 [15], and REAP1 [16].…”

mentioning

confidence: 99%

Mass spectrometric analysis of the editosome and other multiprotein complexes in Trypanosoma brucei

Panigrahi

Allen

Stuart

et al. 2003

J. Am. Soc. Mass Spectrom.

View full text Add to dashboard Cite

The composition of the editosome, a multi-protein complex that catalyzes uridine insertion and deletion RNA editing to produce mature mitochondrial mRNAs in trypanosomes, was analyzed by mass spectrometry. The editosomes were isolated by column chromatography, glycerol gradient sedimentation, and monoclonal antibody affinity purifications. At least 16 proteins form the catalytic core of the editosome, and additional associated proteins were identified. Analyses of mitochondrial fractions identified several non-editosome proteins and multi-protein complexes. These studies contribute to the functional annotation of T. brucei genome. (J Am Soc Mass Spectrom 2003, 14, 728 -735)

show abstract

Cloning and characterization of two guide RNA-binding proteins from mitochondria of Crithidia fasciculata: gBP27, a novel protein, and gBP29, the orthologue of Trypanosoma brucei gBP21

Cited by 51 publications

References 63 publications

A 100-kD complex of two RNA-binding proteins from mitochondria of Leishmania tarentolae catalyzes RNA annealing and interacts with several RNA editing components

A 100-kD complex of two RNA-binding proteins from mitochondria of Leishmania tarentolae catalyzes RNA annealing and interacts with several RNA editing components

Functional complementation of Trypanosoma brucei RNA in vitro editing with recombinant RNA ligase

Mass spectrometric analysis of the editosome and other multiprotein complexes in Trypanosoma brucei

Contact Info

Product

Resources

About