Both sense and antisense RNAs are targets for the sense transgene-induced posttranscriptional silencing mechanism

被引:24
作者
Van Houdt, H [1 ]
Van Montagu, M [1 ]
Depicker, A [1 ]
机构
[1] State Univ Ghent VIB, Dept Plantengenet, Vakgrp Mol Genet, B-9000 Ghent, Belgium
来源
MOLECULAR AND GENERAL GENETICS | 2000年 / 263卷 / 06期
关键词
complementary RNA; cosuppression; RNA turnover; transgenic plant; transient expression;
D O I
10.1007/PL00008700
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
Two stable transgenic tobacco lines were obtained as segregants from a primary transformant. Plants homozygous for a T-DNA inverted repeat locus (HOlo1) showed posttranscriptional gene silencing (PTGS) of the neomycin phosphotransferase II (nptII) transgenes, whereas HOlo2 plants, homozygous for a single T-DNA insert, expressed the nptII genes normally. Transient expression of nptII genes newly introduced into leaves of both the HOlo2 and nptII-silenced HOlo1 plants was downregulated only in the silenced background. Different chimeric beta-glucuronidase (gus) genes with parts of the nptII transgene inserted in sense or antisense orientation into the 3'-untranslated region, which encoded transcripts that had homology or complementarity to nptII transcripts, showed reduced transient expression specifically in npII-silenced tissue. Therefore, we conclude that RNAs of both polarities are targets for PTGS-induced RNA degradation, which supports the notion that double-stranded RNA acts as an inducing signal for silencing.
引用
收藏
页码:995 / 1002
页数:8
相关论文
共 42 条
[1]   Double-stranded RNA specifically disrupts gene expression during planarian regeneration [J].
Alvarado, AS ;
Newmark, PA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (09) :5049-5054
[2]   EXPRESSION OF DICISTRONIC TRANSCRIPTIONAL UNITS IN TRANSGENIC TOBACCO [J].
ANGENON, G ;
UOTILA, J ;
KURKELA, SA ;
TEERI, TH ;
BOTTERMAN, J ;
VANMONTAGU, M ;
DEPICKER, A .
MOLECULAR AND CELLULAR BIOLOGY, 1989, 9 (12) :5676-5684
[3]  
Baulcombe DC, 1996, PLANT CELL, V8, P1833, DOI 10.1105/tpc.8.10.1833
[4]  
Breyne P., 1993, Plant Molecular Biology Reporter, V11, P21, DOI 10.1007/BF02670556
[5]   Plant gene silencing regularized [J].
Bruening, G .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (23) :13349-13351
[6]   Gene silencing in Neurospora crassa requires a protein homologous to RNA-dependent RNA polymerase [J].
Cogoni, C ;
Macino, G .
NATURE, 1999, 399 (6732) :166-169
[7]   Gene silencing results in instability of antibody production in transgenic plants [J].
De Neve M. ;
De Buck S. ;
De Wilde C. ;
Van Houdt H. ;
Strobbe I. ;
Jacobs A. ;
Van Montagu M. ;
Depicker A. .
Molecular and General Genetics MGG, 1999, 260 (6) :582-592
[8]   DIFFERENT 5'-LEADER SEQUENCES MODULATE BETA-GLUCURONIDASE ACCUMULATION LEVELS IN TRANSGENIC NICOTIANA-TABACUM PLANTS [J].
DELOOSE, M ;
DANTHINNE, X ;
VANBOCKSTAELE, E ;
VANMONTAGU, M ;
DEPICKER, A .
EUPHYTICA, 1995, 85 (1-3) :209-216
[9]   Post-transcriptional gene silencing in plants [J].
Depicker, A ;
VanMontagu, M .
CURRENT OPINION IN CELL BIOLOGY, 1997, 9 (03) :373-382
[10]  
Depicker A., 1996, MECH APPL GENE SILEN, P71