Cleavage site mapping and substrate-specificity of Leishmaniavirus 2-1 capsid endoribonuclease activity.
The Leishmaniavirus capsid protein possesses an RNA endoribonuclease activity that cleaves viral positive-sense RNA at a specific, single site within the 5' untranslated region. The site of cleavage in LRV1-4 RNA was previously mapped to nucleotide 320 of the LRV1-4 genome. Here we show that an LRV2-1-derived substrate RNA transcript is also cleaved at a single site in an in vitro cleavage assay with LRV2-1 virions. Precise RNA cleavage site mapping in this divergent Old World virus, LRV2-1, confirms that cleavage is occurring within a region of homology to the LRV1 isolates. Substrate RNA transcripts possessing viral sequences from LRV1-4 or LRV2-1 genomes were assayed for susceptibility to cleavage by the cognate and noncognate capsid endoribonucleases to determine the level of substrate specificity.
MacBeth
K J
KJ
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.
Ro
Y T
YT
Gehrke
L
L
Patterson
J L
JL
D001665:Binding Sites; D002213:Capsid / Q000378:metabolism*; D004722:Endoribonucleases / Q000378:metabolism*; D019199:Leishmaniavirus / Q000737:chemistry*; D014176:Protein Biosynthesis; D012367:RNA, Viral / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity; D014771:Virion / Q000378:metabolism
D012367:RNA, Viral; D004722:Endoribonucleases