Human rotaviruses, which are placed into two groups according to their ribonucleic acid patterns obtained by gel electrophoresis, were characterized both by polypeptide components from purified virions and by polypeptides translated from their denatured ribonucleic acids in rabbit reticulocyte lysates. Viruses assigned to different groups differed in the electrophoretic migration of the second largest of the polypeptides which compose the inner shell; polypeptides that had been synthetized in vitro from ribonucleic acid from each group showed this same difference, thus indicating that this is due to the genomic composition. This study suggests that there are differences in the third largest polypeptide of the inner shell and also in the three smaller polypeptides composing the outer shell. We also demonstrated that there are differences in genomic and polypeptide compositions between simian (SA11) and calf (Nebraska calf diarrhea virus) rotaviruses grown in tissue culture and human rotaviruses.