Rho H M, Green M
Proc Natl Acad Sci U S A. 1974 Jun;71(6):2386-90. doi: 10.1073/pnas.71.6.2386.
Adenosine is the major 3'OH-terminal nucleoside of the 60-70S RNA genome of the murine sarcoma-leukemia virus, its 30-40S RNA subunits, and the poly(A) segments derived by RNase treatment of both RNA species, as determined by periodate oxidation-[(3)H]-borohydride reduction. The binding 30-40S RNA to oligo(dT)-cellulose suggests that most viral RNA subunits contain poly(A). The molecular weight of poly(A) derived from viral RNA by digestion with RNase and purified by affinity chromatography is 64,000-68,000, as determined by gel electrophoresis. From the size of poly(A) and the poly(A) content of viral RNA (1.6%), it is estimated that there is about one poly(A) segment for each viral 30-40S RNA subunit. The results of 3'-termini labeling with [(3)H]borohydride, in vivo labeling with [(3)H]adenosine, and base composition of [(32)P]poly(A) indicate that a homopoly(A) segment is located at the 3'-end of a 30-40S RNA subunit. The homogeneous poly(A) segments isolated from RNase T1 digests of 60-70S [(32)P]RNA consist of one cytidylate, one uridylate, and about 190 adenylate residues, while those isolated from RNase A digests consist exclusively of adenylate residues. These results indicate that -G(C,U)A(190)A(OH) is the 3'-terminal nucleotide sequence of the viral 30-40S RNA subunits.
通过高碘酸盐氧化-[(3)H]-硼氢化钠还原法测定,腺苷是鼠肉瘤-白血病病毒60-70S RNA基因组、其30-40S RNA亚基以及通过对这两种RNA进行核糖核酸酶处理得到的聚(A)片段的主要3'OH末端核苷。30-40S RNA与寡聚(dT)-纤维素的结合表明,大多数病毒RNA亚基含有聚(A)。通过凝胶电泳测定,用核糖核酸酶消化病毒RNA并通过亲和层析纯化得到的聚(A)的分子量为64,000-68,000。根据聚(A)的大小和病毒RNA的聚(A)含量(1.6%)估计,每个病毒30-40S RNA亚基大约有一个聚(A)片段。用[(3)H]硼氢化钠进行3'末端标记、用[(3)H]腺苷进行体内标记以及[(32)P]聚(A)的碱基组成的结果表明,一个同聚(A)片段位于30-40S RNA亚基的3'末端。从60-70S [(32)P]RNA的核糖核酸酶T1消化物中分离出的均匀聚(A)片段由一个胞苷酸、一个尿苷酸和约190个腺苷酸残基组成,而从核糖核酸酶A消化物中分离出的片段仅由腺苷酸残基组成。这些结果表明,-G(C,U)A(190)A(OH)是病毒30-40S RNA亚基的3'末端核苷酸序列。