Disruption of a gene encoding a novel mitochondrial DEAD-box protein in Trypanosoma brucei affects edited mRNAs

被引:97
作者
Missel, A [1 ]
Souza, AE [1 ]
Norskau, G [1 ]
Goringer, HU [1 ]
机构
[1] UNIV MUNICH, GENZENTRUM, MOL BIOL LAB, D-82152 MARTINSRIED, GERMANY
关键词
D O I
10.1128/MCB.17.9.4895
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The majority of mitochondrial pre-mRNAs in kinetoplastid protozoa such as Trypanosoma, Leishmania, and Crithidia are substrates of a posttranscriptional processing reaction referred to as RNA editing. The process results in the insertion and, to a lesser extent, deletion of uridylates, thereby completing the informational content of the mRNAs, The specificity of the RNA editing reaction is provided by guide RNAs (gRNAs), which serve as templates for the editing apparatus, In addition, the process relies on mitochondrial proteins, presumably acting within a high-molecular-mass ribonucleoprotein complex. Although several enzymatic activities have been implicated in the editing process, no protein has been identified to date, Here we report the identification of a novel mitochondrial DEAD-box protein, which we termed mHel61p, Disruption of the mHEL61 alleles in insect-stage Trypanosoma brucei cells resulted in a reduced growth rate phenotype. On a molecular level, the null mutant showed significantly reduced amounts of edited mRNAs, whereas never-edited and nuclear mRNAs were unaffected, Reexpression of mHel61p in the knockout cell line restored the ability to efficiently synthesize edited mRNAs, The results suggest an involvement of mHel61p in the control of the abundance of edited mRNAs and thus reveal a novel function for DEAD-box proteins.
引用
收藏
页码:4895 / 4903
页数:9
相关论文
共 51 条
[1]   MAJOR TRANSCRIPT OF THE FRAMESHIFTED COXLL GENE FROM TRYPANOSOME MITOCHONDRIA CONTAINS 4 NUCLEOTIDES THAT ARE NOT ENCODED IN THE DNA [J].
BENNE, R ;
VANDENBURG, J ;
BRAKENHOFF, JPJ ;
SLOOF, P ;
VANBOOM, JH ;
TROMP, MC .
CELL, 1986, 46 (06) :819-826
[2]  
Beverley S. M., 1993, PROTOCOLS MOL PARASI, P333
[3]   Vectors for inducible expression of toxic gene products in bloodstream and procyclic Trypanosoma brucei [J].
Biebinger, S ;
Wirtz, LE ;
Lorenz, P ;
Clayton, C .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1997, 85 (01) :99-112
[4]   CHARACTERIZATION OF 2 NUCLEAR-ENCODED PROTEIN-COMPONENTS OF MITOCHONDRIAL RIBONUCLEOPROTEIN COMPLEXES FROM LEISHMANIA-TARENTOLAE [J].
BRINGAUD, F ;
PERIS, M ;
ZEN, KH ;
SIMPSON, L .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1995, 71 (01) :65-79
[5]  
BRUN R, 1979, ACTA TROP, V36, P289
[6]   Guide RNA-directed uridine insertion RNA editing in vitro [J].
Byrne, EM ;
Connell, GJ ;
Simpson, L .
EMBO JOURNAL, 1996, 15 (23) :6758-6765
[7]   SINGLE-STEP METHOD OF RNA ISOLATION BY ACID GUANIDINIUM THIOCYANATE PHENOL CHLOROFORM EXTRACTION [J].
CHOMCZYNSKI, P ;
SACCHI, N .
ANALYTICAL BIOCHEMISTRY, 1987, 162 (01) :156-159
[8]  
COMPANY M, 1991, NATURE, V349, P487, DOI 10.1038/349487a0
[9]  
Corell RA, 1996, MOL CELL BIOL, V16, P1410
[10]   IDENTIFICATION, PURIFICATION AND PROPERTIES OF CLONE-SPECIFIC GLYCOPROTEIN ANTIGENS CONSTITUTING SURFACE COAT OF TRYPANOSOMA-BRUCEI [J].
CROSS, GAM .
PARASITOLOGY, 1975, 71 (DEC) :393-+