Nucleic Acids Research, 1981, Vol. 9, No. 7 1571-1581
© 1981
MOLECULAR BIOLOGY |
Ribosomal RNA cleavage, nuclease activation and 2-5A (ppp(A2'p)nA) in interferon-treated cells
National Institute for Medical Research London, NW7 1AA, UK
Received February 5, 1981.
Ribosomal RNA (rRNA) in intact ribosomes is cleaved into discrete products on incubation of reticulocyte lysates or L-cell extracts with ppp(A2'p)3A.Cleavage of rRNA may, therefore, provide a useful assay for 2-5A (ppp(A2'p)n A; n = 2 to 4) or for the presence of a 2-5A-dependent nuclease. The results with reticulocyte lysates differed from those obtained in the L-cell-free system in that (a) a different RNA cleavage pattern was produced (with added L-cell ribosomes) and (b) cleavage was fully activated by the analogue ppp(A2'p)3A3'pCp. As might be expected from the relatively high levels of 2-5A present in interferon-treated, encephalomyocarditis virus (EMC)-infected L-cells, rRNA extracted from these cells was also cleaved. The cleavage pattern observed overlapped with that obtained on incubation of an L-cell-free system with 2-5A. Thus, not only is 2-5A present, but the 2-5A-dependent nuclease also appears to be active, in interferon-treated, EMC-infected L-cells.
1D.H.W. Department of Microbiology, Tel Aviv University, Ramat Aviv, Israel. R.H.S. and I.M.K., ICRF Laboratories, Lincoln's Inn Fields, London, WC2A 3PX.
2D.H.W. Department of Microbiology, Tel Aviv University, Ramat Aviv, Israel. R.H.S. and I.M.K., ICRF Laboratories, Lincoln's Inn Fields, London, WC2A 3PX.
3The 2-5A-dependent nuclease in rabbit reticulocyte lysates is unusual in requiring the tetramer or higher oligomers rather than the trimer or higher oligomers of 2-5A for activity (11)
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