Trans-cleaving hammerhead ribozymes with tertiary stabilizing motifs: in vitro and in vivo activity against a structured viroid RNA

被引：23

作者：

Carbonell, Alberto ^{[1
]}

Flores, Ricardo ^{[1
]}

Gago, Selma ^{[1
]}

机构：

[1] Inst Biol Mol & Celular Plantas UPV CSIC, Valencia 46022, Spain

来源：

NUCLEIC ACIDS RESEARCH | 2011年 / 39卷 / 06期

关键词：

SELF-CLEAVAGE; CATALYTIC CORE; SEQUENCE; VIRUS; REPLICATION; NUCLEOTIDE; INFECTION; KINETICS; INCREASE; PLUS;

D O I：

10.1093/nar/gkq1051

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

070307 [化学生物学]; 071010 [生物化学与分子生物学];

摘要：

Trans-cleaving hammerheads with discontinuous or extended stem I and with tertiary stabilizing motifs (TSMs) have been tested previously against short RNA substrates in vitro at low Mg2+ concentration. However, the potential of these ribozymes for targeting longer and structured RNAs in vitro and in vivo has not been examined. Here, we report the in vitro cleavage of short RNAs and of a 464-nt highly structured RNA from potato spindle tuber viroid (PSTVd) by hammerheads with discontinuous and extended formats at submillimolar Mg2+. Under these conditions, hammerheads derived from eggplant latent viroid and peach latent mosaic viroid (PLMVd) with discontinuous and extended formats, respectively, where the most active. Furthermore, a PLMVd-derived hammerhead with natural TSMs showed activity in vivo against the same long substrate and interfered with systemic PSTVd infection, thus reinforcing the idea that this class of ribozymes has potential to control pathogenic RNA replicons.

引用

页码：2432 / 2444

页数：13

共 43 条

[1]

[Anonymous], 1989, Molecular Cloning: A Laboratory

[2]

EVIDENCE FOR A SINGLE ROLLING CIRCLE IN THE REPLICATION OF POTATO SPINDLE TUBER VIROID [J].

BRANCH, AD ;

BENENFELD, BJ ;

ROBERTSON, HD .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (23) :9128-9132

[3]

Burke Donald H., 2005, BMC Biochemistry, V6, DOI 10.1186/1471-2091-6-14

[4]

The effect of structure in a long target RNA on ribozyme cleavage efficiency [J].

Campbell, TB ;

McDonald, CK ;

Hagen, M .

NUCLEIC ACIDS RESEARCH, 1997, 25 (24) :4985-4993

[5]

Fast cleavage kinetics of a natural hammerhead ribozyme [J].

Canny, MD ;

Jucker, FM ;

Kellogg, E ;

Khvorova, A ;

Jayasena, SD ;

Pardi, A .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2004, 126 (35) :10848-10849

[6]

Double-stranded RNA interferes in a sequence-specific manner with the infection of representative members of the two viroid families [J].

Carbonell, Alberto ;

Martinez de Alba, Angel-Emillo ;

Flores, Ricardo ;

Gago, Selma .

VIROLOGY, 2008, 371 (01) :44-53

[7]

Effects of the trinucleotide preceding the self-cleavage site on eggplant latent viroid hammerheads:: differences in co- and post-transcriptional self-cleavage may explain the lack of trinucleotide AUC in most natural hammerheads [J].

Carbonell, Alberto ;

De la Pena, Marcos ;

Flores, Ricardo ;

Gago, Selma .

NUCLEIC ACIDS RESEARCH, 2006, 34 (19) :5613-5622

[8]

Intracellular ribozyme applications [J].

Castanotto, D ;

Li, JR ;

Michienzi, A ;

Langlois, MA ;

Lee, NS ;

Puymirat, J ;

Rossi, JJ .

BIOCHEMICAL SOCIETY TRANSACTIONS, 2002, 30 :1140-1145

[9]

Capturing hammerhead ribozyme structures in action by modulating general base catalysis [J].

Chi, Young-In ;

Martick, Monika ;

Lares, Monica ;

Kim, Rosalind ;

Scott, William G. ;

Kim, Sung-Hou .

PLOS BIOLOGY, 2008, 6 (09) :2060-2068

[10]

REPLICATION OF AVOCADO SUNBLOTCH VIROID - EVIDENCE FOR A SYMMETRICAL PATHWAY WITH 2 ROLLING CIRCLES AND HAMMERHEAD RIBOZYME PROCESSING [J].

DAROS, JA ;

MARCOS, JF ;

HERNANDEZ, C ;

FLORES, R .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (26) :12813-12817

← 1 2 3 4 5 →