JBC, Vol. 250, Issue 2, 409-417, Jan, 1975

The characteristics of inhibition of protein synthesis by double-stranded ribonucleic acid in reticulocyte lysates

T. Hunter, T. Hunt, R. J. Jackson and H. D. Robertson

All types of double-stranded RNA (DSRNA) tested inhibit protein synthesis in rabbit reticulocyte lysates. The inhibition is characterized by its strongly biphasic kinetics, and can be enhanced by preincubation of the lysate with dsRNA in the absence of protein synthesis. Only properly and extensively matched dsRNA (greater than about 50 base pairs) has this property; no form of DNA, single-stranded RNA or even RNA-DNA hybrids act as inhibitors in this way. The cause of the inhibition appears to be a failure of initiator tRNA to associate with native ribosomal subunits in the initiation process (Darnbrough, C., Hunt, T., and Jackson, R. J. (1973) Biochem. Biophys. Res. Commun. 48, 1556-1564). We have shown that this block is not accompanied by stable association of dsRNA with the ribosomes. There are several reasons to believe that the mechanism of action of dsRNA may be complex with the possible involvement of at least one catalytic step. First, the lysate is inhibited by levels of dsRNA at which ribosomes are present in 100-fold excess over base pairs of dsRNA present. Second, high concentrations of dsRNA (greater than 10 mug per ml) are not inhibitory, but can in some, but not all experiments, reverse the inhibition caused by lower levels of dsRNA. Third, a lysate which has been inhibited by dsRNA, when mixed with a fresh lysate will inhibit synthesis in the mixture much more severely than would be expected from the concentration of dsRNA now present. These results indicate that low levels of dsRNA promote the formation of an inhibitor which may exist in two forms: one that is reversible by high levels of dsRNA and one that is irreversible.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				S. R. Nallagatla and P. C. Bevilacqua Nucleoside modifications modulate activation of the protein kinase PKR in an RNA structure-specific manner RNA, June 1, 2008; 14(6): 1201 - 1213. [Abstract] [Full Text] [PDF]

				A. K. LeFebvre, N. L. Korneeva, M. Trutschl, U. Cvek, R. D. Duzan, C. A. Bradley, J. W. B. Hershey, and R. E. Rhoads Translation Initiation Factor eIF4G-1 Binds to eIF3 through the eIF3e Subunit J. Biol. Chem., August 11, 2006; 281(32): 22917 - 22932. [Abstract] [Full Text] [PDF]

				G. Dotti, B. Savoldo, M. Pule, K. C. Straathof, E. Biagi, E. Yvon, S. Vigouroux, M. K. Brenner, and C. M. Rooney Human cytotoxic T lymphocytes with reduced sensitivity to Fas-induced apoptosis Blood, June 15, 2005; 105(12): 4677 - 4684. [Abstract] [Full Text] [PDF]

				S. Lu, R. Shi, C.-C. Tsao, X. Yi, L. Li, and V. L. Chiang RNA silencing in plants by the expression of siRNA duplexes Nucleic Acids Res., December 2, 2004; 32(21): e171 - e171. [Abstract] [Full Text] [PDF]

				S. Stier, T. Cheng, R. Forkert, C. Lutz, D. M. Dombkowski, J. L. Zhang, and D. T. Scadden Ex vivo targeting of p21Cip1/Waf1 permits relative expansion of human hematopoietic stem cells Blood, August 15, 2003; 102(4): 1260 - 1266. [Abstract] [Full Text] [PDF]

				L. R. SAUNDERS and G. N. BARBER The dsRNA binding protein family: critical roles, diverse cellular functions FASEB J, June 1, 2003; 17(9): 961 - 983. [Abstract] [Full Text] [PDF]

				J. Wang, E. Tekle, H. Oubrahim, J. J. Mieyal, E. R. Stadtman, and P. B. Chock Stable and controllable RNA interference: Investigating the physiological function of glutathionylated actin PNAS, April 29, 2003; 100(9): 5103 - 5106. [Abstract] [Full Text] [PDF]

				G. Tiscornia, O. Singer, M. Ikawa, and I. M. Verma A general method for gene knockdown in mice by using lentiviral vectors expressing small interfering RNA PNAS, February 18, 2003; 100(4): 1844 - 1848. [Abstract] [Full Text] [PDF]

				J. Martínez-Costas, C. González-López, V. N. Vakharia, and J. Benavente Possible Involvement of the Double-Stranded RNA-Binding Core Protein sigma A in the Resistance of Avian Reovirus to Interferon J. Virol., February 1, 2000; 74(3): 1124 - 1131. [Abstract] [Full Text]

				H. K. Tirupati, L. C. Shaw, and A. S. Lewin An RNA Binding Motif in the Cbp2 Protein Required for Protein-stimulated RNA Catalysis J. Biol. Chem., October 22, 1999; 274(43): 30393 - 30401. [Abstract] [Full Text] [PDF]

				K. U. Kumar, S. P. Srivastava, and R. J. Kaufman Double-Stranded RNA-Activated Protein Kinase (PKR) Is Negatively Regulated by 60S Ribosomal Subunit Protein L18 Mol. Cell. Biol., February 1, 1999; 19(2): 1116 - 1125. [Abstract] [Full Text] [PDF]

				P. R. Romano, F. Zhang, S.-L. Tan, M. T. Garcia-Barrio, M. G. Katze, T. E. Dever, and A. G. Hinnebusch Inhibition of Double-Stranded RNA-Dependent Protein Kinase PKR by Vaccinia Virus E3: Role of Complex Formation and the E3 N-Terminal Domain Mol. Cell. Biol., December 1, 1998; 18(12): 7304 - 7316. [Abstract] [Full Text]

				S R Green and M B Mathews Two RNA-binding motifs in the double-stranded RNA-activated protein kinase, DAI. Genes & Dev., December 1, 1992; 6(12b): 2478 - 2490. [Abstract] [PDF]

				H. Hiddinga, C. Crum, J Hu, and D. Roth Viroid-induced phosphorylation of a host protein related to a dsRNA-dependent protein kinase Science, July 22, 1988; 241(4864): 451 - 453. [Abstract] [PDF]

				T. R. Brummelkamp, R. Bernards, and R. Agami A System for Stable Expression of Short Interfering RNAs in Mammalian Cells Science, April 19, 2002; 296(5567): 550 - 553. [Abstract] [Full Text] [PDF]

				F. Zhang, P. R. Romano, T. Nagamura-Inoue, B. Tian, T. E. Dever, M. B. Mathews, K. Ozato, and A. G. Hinnebusch Binding of Double-stranded RNA to Protein Kinase PKR Is Required for Dimerization and Promotes Critical Autophosphorylation Events in the Activation Loop J. Biol. Chem., June 29, 2001; 276(27): 24946 - 24958. [Abstract] [Full Text] [PDF]