Silencing the Drosophila ribosomal protein L14 gene using targeted RNA interference causes distinct somatic anomalies

被引:20
作者
Enerly, E
Larsson, J
Lambertsson, A
机构
[1] Univ Oslo, Div Cell & Mol Biol, Inst Biol, N-0315 Oslo, Norway
[2] Umea Univ, UCMP, SE-90187 Umea, Sweden
关键词
development; gene knockout; ribosome; RNAi; RpL14;
D O I
10.1016/S0378-1119(03)00827-8
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
The Drosophila Minutes are haploinsufficient mutations that are defective in ribosomal protein (rp) production, resulting in short, thin bristles, delayed development and recessive lethality. In a Minute fly, the amount of rp gene messenger RNA (mRNA) is reduced to greater than or equal to 50% of the normal amount of gene product, and becomes rate limiting for ribosome biogenesis, cell proliferation and growth. Haploinsufficiency increases the vulnerability to complete loss of gene function (homozygous null state) if hit by a second mutation. Because of the homozygous lethality, it has only been possible to study the effects of Minute mutations in heterozygous animals. To be able to study the consequences of a loss-of-function of an rp gene (0% > mRNA < 50%) in developing and differentiated cells we used heritable RNA interference (RNAi) in combination with the yeast GAL4/UAS binary system to spatiotemporally knock down the ribosomal protein L14 (RpL14) gene. We show, at the RNA and phenotypic levels, that RNAi efficiently reduces RpL14 gene expression throughout development, causing lethality and distinct and dramatic somatic anomalies in both developing and differentiated cells. (C) 2003 Elsevier B.V. All rights reserved.
引用
收藏
页码:41 / 48
页数:8
相关论文
共 38 条
[1]   The genome sequence of Drosophila melanogaster [J].
Adams, MD ;
Celniker, SE ;
Holt, RA ;
Evans, CA ;
Gocayne, JD ;
Amanatides, PG ;
Scherer, SE ;
Li, PW ;
Hoskins, RA ;
Galle, RF ;
George, RA ;
Lewis, SE ;
Richards, S ;
Ashburner, M ;
Henderson, SN ;
Sutton, GG ;
Wortman, JR ;
Yandell, MD ;
Zhang, Q ;
Chen, LX ;
Brandon, RC ;
Rogers, YHC ;
Blazej, RG ;
Champe, M ;
Pfeiffer, BD ;
Wan, KH ;
Doyle, C ;
Baxter, EG ;
Helt, G ;
Nelson, CR ;
Miklos, GLG ;
Abril, JF ;
Agbayani, A ;
An, HJ ;
Andrews-Pfannkoch, C ;
Baldwin, D ;
Ballew, RM ;
Basu, A ;
Baxendale, J ;
Bayraktaroglu, L ;
Beasley, EM ;
Beeson, KY ;
Benos, PV ;
Berman, BP ;
Bhandari, D ;
Bolshakov, S ;
Borkova, D ;
Botchan, MR ;
Bouck, J ;
Brokstein, P .
SCIENCE, 2000, 287 (5461) :2185-2195
[2]   Dosage-dependent gene regulation in multicellular eukaryotes: Implications for dosage compensation, aneuploid syndromes, and quantitative traits [J].
Birchler, JA ;
Bhadra, U ;
Bhadra, MP ;
Auger, DL .
DEVELOPMENTAL BIOLOGY, 2001, 234 (02) :275-288
[3]   Differential expression of genes coding for ribosomal proteins in different human tissues [J].
Bortoluzzi, S ;
d'Alessi, F ;
Romualdi, C ;
Danieli, GA .
BIOINFORMATICS, 2001, 17 (12) :1152-1157
[4]  
BRAND AH, 1993, DEVELOPMENT, V118, P401
[5]   Visualization of gene expression in living adult Drosophila [J].
Calleja, M ;
Moreno, E ;
Pelaz, S ;
Morata, G .
SCIENCE, 1996, 274 (5285) :252-255
[6]   Associative learning disrupted by impaired G(s) signaling in Drosophila mushroom bodies [J].
Connolly, JB ;
Roberts, IJH ;
Armstrong, JD ;
Kaiser, K ;
Forte, M ;
Tully, T ;
OKane, CJ .
SCIENCE, 1996, 274 (5295) :2104-2107
[7]   Modeling stochastic gene expression: Implications for haploinsufficiency [J].
Cook, DL ;
Gerber, LN ;
Tapscott, SJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (26) :15641-15646
[8]  
CRAMTON SE, 1994, GENETICS, V137, P1039
[9]   The gene encoding ribosomal protein S19 is mutated in Diamond-Blackfan anaemia [J].
Draptchinskaia, N ;
Gustavsson, P ;
Andersson, B ;
Pettersson, M ;
Willig, TN ;
Dianzani, I ;
Ball, S ;
Tchernia, G ;
Klar, J ;
Matsson, H ;
Tentler, D ;
Mohandas, N ;
Carlsson, B ;
Dahl, N .
NATURE GENETICS, 1999, 21 (02) :169-175
[10]   Reverse genetics in Drosophila:: From sequence to phenotype using UAS-RNAi transgenic flies [J].
Enerly, E ;
Larsson, J ;
Lambertsson, A .
GENESIS, 2002, 34 (1-2) :152-155