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Published October 28, 1969 | public
Journal Article

The process of infection with bacteriophage φX174: XXIX. In vivo studies on the synthesis of the single-stranded DNA of progeny φX174 bacteriophage

Abstract

Double-stranded φXDNA rings, composed of a closed complementary strand and an open viral strand, are found in φX-infected cells during the period of progeny single-stranded DNA formation. The linear viral strand components of these double-stranded circles are nicked between a deoxyguanosine (on the 3′-terminus) and a deoxycytidine (on the 5′-terminus). The 5′-terminus is not phosphorylated. Upon replication, nucleotides are added to the 3′-end of the viral strand. The 5′ end is concomitantly displaced from its complementary strand template and appears, by electron microscopy and by column chromatography, as a single-stranded, non-hydrogen-bonded, tail of a double-stranded circle. While the viral strand grows (to a maximum of twice the length of one φX genome) by addition of nucleotides to the 3′-end, an interesting reaction, of uncertain nature, also takes place at the free 5′-end of the nascent viral strand, the net result of which is that the original terminal deoxycytidine is replaced by a terminal deoxyguanosine. Our experiments indicate that the single-stranded DNA synthesis does not take place at the cell membrane where double-stranded φX DNA is formed but upon "cytoplasmic" RF‡. Viral coat protein can be shown to be associated with the single-strand-producing complexes.

Additional Information

© 1969 Elsevier Ltd. Received 28 February 1969, Revised 3 July 1969. This research was supported in part by a grant, GM13554 from the U.S. Public Health Service and a postdoctoral research fellowship 1F05, TW-1348-01 to one of us (A. R.). Robert Rohwer performed the radio immunoassay of the experiment in Figure 8(b).

Additional details

Created:
August 19, 2023
Modified:
October 20, 2023