Molecular evolution of bacteriophages: evidence of selection against the recognition sites of host restriction enzymes

Restriction enzymes produced by bacteria serve as a defense against invading bacteriophages, and so phages without other protection would be expected to undergo selection to eliminate recognition sites for these enzymes from their genomes. The observed frequencies of all restriction sites in the genomes of all completely sequenced DNA phages (T7, lambda, phi X174, G4, M13, f1, fd, and IKe) have been compared to expected frequencies derived from trinucleotide frequencies. Attention was focused on 6-base palindromes since they comprise the typical recognition sites for type II restriction enzymes. All of these coliphages, with the exception of lambda and G4, exhibit significant avoidance of the particular sequences that are enterobacterial restriction sites. As expected, the sequenced fraction of the genome of phi 29, a Bacillus subtilis phage, lacks Bacillus restriction sites. By contrast, the RNA phage MS2, several viruses that infect eukaryotes (EBV, adenovirus, papilloma, and SV40), and three mitochondrial genomes (human, mouse, and cow) were found not to lack restriction sites. Because the particular palindromes avoided correspond closely with the recognition sites for host enzymes and because other viruses and small genomes do not show this avoidance, it is concluded that the effect indeed results from natural selection.      

LOCALIZATION OF TRANSPORT ADENOSINE TRIPHOSPHATASE IN RAT CORNEA

Increasingly, physiological and biochemical data on the mammalian cornea have emphasized the importance of the endothelial rather than the epithelial layer in regulating the state of hydration of the cornea, via a transport ATPase. This view is supported by the observations reported here. With the Ernst method, the first cytochemical procedure to be readily responsive to ions and ouabain, the major sites of enzyme reaction product are the intercellular spaces of the endothelial layer. This localization was not found by us with the so-called ATPase method of Wachstein and Meisel.