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SUMMARYWe have compared the structural polypeptides of an adult T-cell leukaemia (ATL) virus (ATLV) isolate from a Japanese patient with ATL with those of a similar virus derived from a Japanese macaque monkey. Both are distinct but related entities. Their core polypeptides p19 could not be distinguished, but p24, another core polypeptide, and their envelope glycopolypeptides differ. The human virus directs the synthesis of a single intracellular glycopolypeptide, gp68, while the macaque virus specifies two such glycopolypeptides, gp57 and gp50. Furthermore, the glycopolypeptides of both viruses are serologically distinct. Thus, these viruses represent subtypes of the ATLV family and the macaque virus is apparently not involved in human ATL.

Total polysomal poly(A)+ RNA of Novikoff hepatoma and of 18-hr regenerating rat liver were compared by analysis of their in vitro translational products on two-dimensional isofocusing/sodium dodecyl suffate gels. This technique resolved the translated proteins sufficiently to permit detection of quantitative and some qualitative differences between the two mRNA populations. Excess (5-7); however, it is virtually impossible to detect distinct mRNA species that contribute less than 1% of the total mRNA population. Detection of mRNA species present in small amounts can be achieved when the hybridization of mRNA to solid-phase cDNA, as described by Venetianer and Leder (8), is combined with the analysis of the translational products of the hybridized mRNA. Recent improvements in two-dimensional gel electrophoresis (9, 10) were utilized in these studies to achieve a high resolution of the translated proteins. ,ug of actinomycin D per ml, 1% ethanol, 15 mg of oligo(dT)-cellulose per ml, 25 ,ug of mRNA per ml, and 55 units of reverse transcriptase per ml. After a 5-min incubation on ice, the sample was incubated at 420 for 60 min and agitated in a shaking water bath (70 strokes per min) to suspend the cellulose. [8-3H]dGTP (10 ,Ci/ml) was used to label the cDNA. cDNA synthesis was approximately 45% with respect to input mRNA (14). MATERIALS AND METHODSHybridization of mRNA to Oligo(dT)Cellulose cDNA.Twenty micrograms of mRNA was hybridized to a 10-fold excess of cDNA (approximately 200 ,ug), covalently linked to 0.3 g of cellulose, at 40°for approximately 50 hr in a total volume of 3 ml containing 50% formamide, 0.6 M NaCl, 0.1% sodium dodecyl sulfate, 10 mM Tris-HCl (pH 7.6), 1 mM EDTA, and 0.5 A260 unit of oligo(A) per ml (14,15). After the unbound mRNA was washed off the cellulose, the bound mRNA was eluted with 50% formamide/0. 1% sodium dodecyl sulfate/10 mM pH 7.6, at 60°(14). To the eluted fraction, 0.5 A2W unit of oligo(A) per ml was added. The mRNA of both fractions was precipitated by addition of 2.5 volumes of 100% ethanol saturated with sodium acetate. Bound and unbound mRNA were then translated in the wheat germ cellfree system (16,17)