TY - JOUR
T1 - rRNA cleavage as an index of ppp(A2'p)(n)A activity in interferon-treated encephalomyocarditis virus-infected cells
AU - Silverman, R. H.
AU - Skehel, J. J.
AU - James, T. C.
AU - Wreschner, D. H.
AU - Kerr, I. M.
PY - 1983
Y1 - 1983
N2 - In cell-free systems, 2-5A [ppp(A2'p)(n)A, n = 2 to ≥4] activates a latent endoribonuclease, the 2-5A-dependent RNase, which cleaves rRNA in intact ribosomes into discrete and characteristic products. Here we present Northern blots which have identified the 18S or 28S origins of the cleaved products from rRNA. In addition, identical 3' termini were observed for fragments of 18S rRNA from a HeLa cell-free system incubated with 2-5A and from interferon-treated, encephalomyocarditis virus-infected HeLa cells. The previous assumption of identity of such fragments was based only on comigration on electrophoresis in agarose gels. We conclude that appropriate patterns of cleavage found in RNA isolated from intact cells are an indicator of prior 2-5A-dependent RNase activity. The assay of rRNA cleavage is relatively convenient and unambiguous. Accordingly, in the search for situations in which the 2-5A system may be active, it provides a useful alternative to the direct assay of 2-5A.
AB - In cell-free systems, 2-5A [ppp(A2'p)(n)A, n = 2 to ≥4] activates a latent endoribonuclease, the 2-5A-dependent RNase, which cleaves rRNA in intact ribosomes into discrete and characteristic products. Here we present Northern blots which have identified the 18S or 28S origins of the cleaved products from rRNA. In addition, identical 3' termini were observed for fragments of 18S rRNA from a HeLa cell-free system incubated with 2-5A and from interferon-treated, encephalomyocarditis virus-infected HeLa cells. The previous assumption of identity of such fragments was based only on comigration on electrophoresis in agarose gels. We conclude that appropriate patterns of cleavage found in RNA isolated from intact cells are an indicator of prior 2-5A-dependent RNase activity. The assay of rRNA cleavage is relatively convenient and unambiguous. Accordingly, in the search for situations in which the 2-5A system may be active, it provides a useful alternative to the direct assay of 2-5A.
UR - http://www.scopus.com/inward/record.url?scp=0020535757&partnerID=8YFLogxK
U2 - 10.1128/jvi.46.3.1051-1055.1983
DO - 10.1128/jvi.46.3.1051-1055.1983
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AN - SCOPUS:0020535757
VL - 46
SP - 1051
EP - 1055
JO - Unknown Journal
JF - Unknown Journal
IS - 3
ER -