Ryskov A P, Ivanov P L, Kramerov D A, Georgiev G P
Mol Biol (Mosk). 1984 Jan-Feb;18(1):92-103.
A cDNA library in pBR322 was prepared with cytoplasmic poly(A) + RNA from mouse liver cells. From 1 to 1.5% of clones hybridized to either B1 or B2 ubiquitous repetitive sequences. Several clones hybridizing to a B2 repeat were partially sequenced. The full-length B2 sequence was found at the 3'-end of abundant 20S poly(A) + RNA (designated as B2 + mRNA chi) within the non-coding part of it. B2 + mRNA chi is tissue-specific, concentrated in mouse liver polysomes and absent in cytoplasm of Ehrlich carcinoma cells. The B2 sequence seems to be located at the 3'-end of some other mRNAs as well. To determine the orientation of the B2 sequence in different RNAs, its two strands were labeled, electrophoretically separated, and used for hybridization with Northern blots containing nuclear, cytoplasmic and polysomal RNAs. In nuclear RNA, the B2 sequence is present in both orientations; in polysomal and cytoplasmic poly(A) + RNAs, only one ("canonical") strand of it can be detected. Low molecular weight poly(A) + B2 + RNA also contains the same strand of the B2 element. The conclusion has been drawn that only one of its strand can survive the processing. This strand contains promoter-like sequences and AATAAA blocks. The latter can be used in some cases by the cell as mRNA polyadenylation signals.
用小鼠肝细胞的细胞质多聚腺苷酸(poly(A))+RNA构建了pBR322中的cDNA文库。1%至1.5%的克隆与B1或B2普遍存在的重复序列杂交。对几个与B2重复序列杂交的克隆进行了部分测序。在丰富的20S多聚腺苷酸(poly(A))+RNA(命名为B2+mRNA chi)的非编码区3'端发现了完整的B2序列。B2+mRNA chi具有组织特异性,集中在小鼠肝脏多核糖体中,而艾氏癌细胞的细胞质中不存在。B2序列似乎也位于其他一些mRNA的3'端。为了确定B2序列在不同RNA中的方向,对其两条链进行标记,通过电泳分离,然后用于与包含核RNA、细胞质RNA和多核糖体RNA的Northern印迹杂交。在核RNA中,B2序列以两种方向存在;在多核糖体和细胞质多聚腺苷酸(poly(A))+RNA中,只能检测到其一条(“标准”)链。低分子量多聚腺苷酸(poly(A))+B2+RNA也包含B2元件的同一条链。得出的结论是,其只有一条链能够在加工过程中留存下来。这条链包含类似启动子的序列和AATAAA序列块。在某些情况下,细胞可以将后者用作mRNA聚腺苷酸化信号。