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The process of infection with bacteriophage φX174. XXXI. Abortive Infection at Low Temperatures

Newbold, John E. and Sinsheimer, Robert L. (1970) The process of infection with bacteriophage φX174. XXXI. Abortive Infection at Low Temperatures. Journal of Molecular Biology, 49 (1). pp. 23-47. ISSN 0022-2836. https://resolver.caltech.edu/CaltechAUTHORS:20160802-150703121

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Abstract

The infection of Escherichia coli by φX174 at 15 °C is abortive; the cells are killed by the infection but neither mature phage nor SS-DNA are synthesized. Parental RF is formed and subsequently replicated at 15 °C. The RF made at 15 °C shows normal infectivity and competence as a precursor to progeny SS-DNA after an increase in temperature to 37 °C. Some 200 phage equivalents of serum blocking power are synthesized also during 12 hours of infection at 15 °C. The addition of chloramphenicol to 100 μg/ml. to abortively infected cells late in the infection at 15 °C does not block the subsequent maturation of infectious virus following an increase in temperature to 37 °C. The data suggest that, given an adequate pool of the essential proteins and the precursor RF, the concomitant synthesis of protein is not required for the synthesis of SS-DNA or for phage maturation. It is concluded that all of the proteins required for SS-DNA synthesis and phage maturation are present in the abortive infection at 15 °C. Temperature “shift-up” experiments (from 15 up to 41 °C) performed with temperature-sensitive mutants of φX174 and carried out in the presence of chloramphenicol as just described further demonstrate that the proteins coded by φX174 cistrons II, III, IV, V and VII are synthesized at 15 °C and probably adopt their native conformation at this low temperature. The proteins coded by several φX174 strains mutant in cistrons II, III, IV and VII are shown to be functional at 41 °C, when synthesized at 15 °C, although they are inactive when synthesized at 41 °C. These experiments indicate that the majority of ts mutations impose their restriction during the folding of the nascent polypeptide into the native conformation. Three possible causes are suggested for the abortive infection at 15 °C. (a) A virus-coded protein whose role is essential to the infection is made at 15 °C and assumes its native conformation but its rate of activity is too slow at this temperature to sustain the infection process. (b) Virus maturation may involve the formation of a DNA-protein complex and conformational changes which have an energy threshold infrequently reached at 15 °C. (c) A host-coded protein present in uninfected cells, and whose activity is essential to the infection at all temperatures but not to the host at 15 °C, is inactive at 15 °C. An hypothesis of this type is offered which proposes that the temperature-limiting factor in SS-DNA synthesis in vivo may reflect a temperature-dependent property of the host DNA polymerase. The low temperature threshold for SS-DNA synthesis and phage maturation in infected cells is close to 17 °C.


Item Type:Article
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http://dx.doi.org/10.1016/0022-2836(70)90374-8DOIArticle
http://www.sciencedirect.com/science/article/pii/0022283670903748PublisherArticle
Additional Information:© 1970 Elsevier Ltd. Received 1 August 1969, Revised 22 December 1969. We would like to acknowledge useful discussions with Clyde A. Hutchison, III during the course of this work. The authors are grateful to Robert Rohwer who performed the serum blocking power assays reported in Table 3 and to Dr Robert Mayol who undertook the electrophoresis analysis mentioned in the Discussion. One of us (J.E.N.) gratefully acknowledges the support of fellowships sponsored by the McCallum Fund and the Woodrow Wilson Foundation. This research was supported in part from Public Health Service grant no. GM13554.
Funders:
Funding AgencyGrant Number
McCallum FundUNSPECIFIED
Woodrow Wilson FoundationUNSPECIFIED
U.S. Public Health Service (USPHS)GM13554
Issue or Number:1
Record Number:CaltechAUTHORS:20160802-150703121
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20160802-150703121
Official Citation:John E. Newbold, Robert L. Sinsheimer, The process of infection with bacteriophage φX174: XXXI. Abortive infection at low temperatures, Journal of Molecular Biology, Volume 49, Issue 1, 14 April 1970, Pages 23-47, ISSN 0022-2836, http://dx.doi.org/10.1016/0022-2836(70)90374-8. (http://www.sciencedirect.com/science/article/pii/0022283670903748)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:69398
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:03 Aug 2016 16:52
Last Modified:03 Oct 2019 10:21

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