Reconstitution of rods from tobacco mosaic virus protein and RNA modified with bulky carcinogens.

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Tobacco mosaic virus (TMV) RNA was treated with radioactive N-acetoxy-2-acetylaminofluorene (N-acetoxy-AAF) and (+/-)-7 beta, 8 alpha-dihydroxy-9 alpha, 10 alpha-epoxy-7,8,9,10-tetrahydrobenzo(a)pyrene (BaP diol epoxide) to obtain 3-25 adducts per molecule. Modified full length 30S RNAs and unmodified RNA were reconstituted for various time periods with TMV protein. The particulate products were separated by ultracentrifugation, and the amounts of virus-like material were quantitated by UV spectrophotometry. The length distribution and general appearance of the virus-like rods were studied by electron microscopy. Neither type of carcinogen prevented typical rod formation, but the rate of formation and the maximal yield of reconstituted particles diminished with increasing modification by both agents. The rod length distribution also showed progressively lesser numbers of full-length virus rods. The particulate material contained approximately the same number of adducts as the modified RNA. Thus, it appears that these carcinogen modifications of guanine residues at the N-2 or C-8 atoms did not prevent orderly protein assembly on the RNA but instead slowed up this process and frequently stopped it, possibly at sites where adducts happen to be clustered.

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