HeLa Cell Cytoplasmic mRNA Contains Three Classes of Sequences Predominantly Poly(A)‐Free, Predominantly Poly(A)‐Containing and Bimorphic

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Abstract
The mRNA species which exist in the HeLa cell polyribosomes in a form devoid of A sequences longer than 8 nucleotides constitute the poly(A)‐free class of mRNA. The rapidly labelled component of this mRNA class shares no measurable sequence homology with poly(A)‐containing RNA. If poly(A)‐free mRNA larger than 12 S labelled for 2 h in vivo is hybridized with total cellular DNA, it hybridizes primarily with single‐copy DNA. When a large excess of steady poly(A)‐containing RNA is added before hybridization of labelled poly(A)‐free RNA, no inhibition of hybridization occurs. This indicates the existence of a class of poly(A)‐free mRNA with no poly(A)‐containing counterpart. Some mRNA species can exist solely as poly(A)‐containing mRNAs. These mRNAs in HeLa cells are found almost exclusively in the mRNA species present only a few times per cell (scarce sequences). Some mRNA species can exist in two forms, poly(A)containing and lacking, as evidenced by the translation data in vitro of Kaufmann et al. [Proc. Natl Acad. Sci. U.S.A. 74, 4801–4805 (1977)]. In addition, if cDNA to total poly(A)‐containing mRNA is fractionated into abundant and scarce classes, 47 % of the scarce class cDNA can be readily hybridized with poly(A)‐free mRNA. 10 of the abundant cDNA to poly(A)‐containing mRNA will hybridize with poly(A)‐free sequences very rapidly while the other 90% hybridize 160 times more slowly, indicating two very different frequency distributions. The cytoplasmic metabolism of these three distinct mRNA classes is discussed.