MECHANISMS OF THE INHIBITION OF REVERSE TRANSCRIPTION BY ANTISENSE OLIGONUCLEOTIDES

被引:70
作者
BOIZIAU, C
THUONG, NT
TOULME, JJ
机构
[1] UNIV BORDEAUX 2,INSERM,BIOPHYS MOLEC LAB,CJF90-13,146 RUE LEO SAIGNAT,F-33076 BORDEAUX,FRANCE
[2] CTR BIOPHYS MOLEC,CNRS,F-45071 ORLEANS 2,FRANCE
关键词
SYNTHETIC OLIGODEOXYNUCLEOTIDES; REVERSE TRANSCRIPTASE; RETROVIRUSES;
D O I
10.1073/pnas.89.2.768
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
We have demonstrated that the synthesis of cDNA by avian myeloblastosis virus and Moloney murine leukemia virus reverse transcriptases can be prevented by oligonucleotides bound to the RNA template almost-equal-to 100 nucleotides remote from the 3' end of the primer. The RNA was truncated at the level of the antisense oligonucleotide-RNA duplex during the reverse transcription. The key role played by the reverse transcriptase-associated RNase H activity in the inhibition process was shown by the use of (i) inhibitors of RNase H (NaF or dAMP), (ii) Moloney murine leukemia virus reverse transcriptase devoid of RNase H activity, or (iii) alpha-analogues of oligomers that do not elicit RNase H-catalyzed RNA degradation. In all three cases the inhibitory effect was either reduced (NaF, dAMP) or totally abolished. However, an alpha-oligomer bound to the sequence immediately adjacent to the primer-binding site prevented reverse transcription. Therefore, initiation of polymerization can be blocked by means of an RNase H-independent mechanism, whereas arrest of a growing cDNA strand can be achieved only by an oligonucleotide mediating cleavage of the template RNA.
引用
收藏
页码:768 / 772
页数:5
相关论文
共 32 条
[1]   REVERSE-TRANSCRIPTASE AND ITS ASSOCIATED RIBONUCLEASE-H - INTERPLAY OF 2 ENZYME-ACTIVITIES CONTROLS THE YIELD OF SINGLE-STRANDED COMPLEMENTARY DEOXYRIBONUCLEIC-ACID [J].
BERGER, SL ;
WALLACE, DM ;
PUSKAS, RS ;
ESCHENFELDT, WH .
BIOCHEMISTRY, 1983, 22 (10) :2365-2372
[2]   ALPHA-ANOMERIC DNA - BETA-RNA HYBRIDS AS NEW SYNTHETIC INHIBITORS OF ESCHERICHIA-COLI RNASE-H, DROSOPHILA EMBRYO RNASE-H AND M-MLV REVERSE-TRANSCRIPTASE [J].
BLOCH, E ;
LAVIGNON, M ;
BERTRAND, JR ;
POGNAN, F ;
MORVAN, F ;
MALVY, C ;
RAYNER, B ;
IMBACH, JL ;
PAOLETTI, C .
GENE, 1988, 72 (1-2) :349-360
[3]   INHIBITION OF TRANSLATION INITIATION BY ANTISENSE OLIGONUCLEOTIDES VIA AN RNASE-H INDEPENDENT MECHANISM [J].
BOIZIAU, C ;
KURFURST, R ;
CAZENAVE, C ;
ROIG, V ;
THUONG, NT ;
TOULME, JJ .
NUCLEIC ACIDS RESEARCH, 1991, 19 (05) :1113-1119
[4]  
BOIZIAU C, 1991, NUCLEIC ACIDS RES S, V24, P121
[5]   MECHANISTIC INDEPENDENCE OF AVIAN-MYELOBLASTOSIS VIRUS DNA-POLYMERASE AND RIBONUCLEASE H [J].
BREWER, LC ;
WELLS, RD .
JOURNAL OF VIROLOGY, 1974, 14 (06) :1494-1502
[6]   RATE OF DEGRADATION OF [ALPHA]-OLIGODEOXYNUCLEOTIDES AND [BETA]-OLIGODEOXYNUCLEOTIDES IN XENOPUS OOCYTES - IMPLICATIONS FOR ANTI-MESSENGER STRATEGIES [J].
CAZENAVE, C ;
CHEVRIER, M ;
THUONG, NT ;
HELENE, C .
NUCLEIC ACIDS RESEARCH, 1987, 15 (24) :10507-10521
[7]   COMPARATIVE INHIBITION OF RABBIT GLOBIN MESSENGER-RNA TRANSLATION BY MODIFIED ANTISENSE OLIGODEOXYNUCLEOTIDES [J].
CAZENAVE, C ;
STEIN, CA ;
LOREAU, N ;
THUONG, NT ;
NECKERS, LM ;
SUBASINGHE, C ;
HELENE, C ;
COHEN, JS ;
TOULME, JJ .
NUCLEIC ACIDS RESEARCH, 1989, 17 (11) :4255-4273
[8]   INHIBITION OF DNA-SYNTHESIS BY CROSS-LINKING THE TEMPLATE TO PLATINUM-THIOL DERIVATIVES OF COMPLEMENTARY OLIGODEOXYNUCLEOTIDES [J].
CHU, BCF ;
ORGEL, LE .
NUCLEIC ACIDS RESEARCH, 1989, 17 (12) :4783-4798
[9]  
DESTEFANO JJ, 1991, J BIOL CHEM, V266, P7423
[10]  
FURFINE ES, 1991, J BIOL CHEM, V266, P406