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Article

Mutagenic Analysis of a DNA Translocating Tube’s Interior Surface

1
The BIO5 Institute, University of Arizona, Tucson, AZ 85721, USA
2
Statistics Consulting Lab, The BIO5 Institute, University of Arizona, Tucson, AZ 85721, USA
*
Author to whom correspondence should be addressed.
Current address: Institute for Cell and Molecular Biology, Department of Molecular Biosciences, University of Texas, Austin, TX 78712, USA
Received: 19 May 2020 / Revised: 17 June 2020 / Accepted: 18 June 2020 / Published: 22 June 2020
(This article belongs to the Special Issue In Memory of Michael Rossmann)
Bacteriophage ϕX174 uses a decamer of DNA piloting proteins to penetrate its host. These proteins oligomerize into a cell wall-spanning tube, wide enough for genome passage. While the inner surface of the tube is primarily lined with inward-facing amino acid side chains containing amide and guanidinium groups, there is a 28 Å-long section near the tube’s C-terminus that does not exhibit this motif. The majority of the inward-facing residues in this region are conserved across the three ϕX174-like clades, suggesting that they play an important role during genome delivery. To test this hypothesis, and explore the general function of the tube’s inner surface, non-glutamine residues within this region were mutated to glutamine, while existing glutamine residues were changed to serine. Four of the resulting mutants had temperature-dependent phenotypes. Virion assembly, host attachment, and virion eclipse, defined as the cell’s ability to inactivate the virus, were not affected. Genome delivery, however, was inhibited. The results support a model in which a balance of forces governs genome delivery: potential energy provided by the densely packaged viral genome and/or an osmotic gradient move the genome into the cell, while the tube’s inward facing glutamine residues exert a frictional force, or drag, that controls genome release. View Full-Text
Keywords: ϕX174; microviridae; penetration; DNA pilot protein ϕX174; microviridae; penetration; DNA pilot protein
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MDPI and ACS Style

Roznowski, A.P.; Fisher, J.M.; Fane, B.A. Mutagenic Analysis of a DNA Translocating Tube’s Interior Surface. Viruses 2020, 12, 670. https://0-doi-org.brum.beds.ac.uk/10.3390/v12060670

AMA Style

Roznowski AP, Fisher JM, Fane BA. Mutagenic Analysis of a DNA Translocating Tube’s Interior Surface. Viruses. 2020; 12(6):670. https://0-doi-org.brum.beds.ac.uk/10.3390/v12060670

Chicago/Turabian Style

Roznowski, Aaron P., Julia M. Fisher, and Bentley A. Fane 2020. "Mutagenic Analysis of a DNA Translocating Tube’s Interior Surface" Viruses 12, no. 6: 670. https://0-doi-org.brum.beds.ac.uk/10.3390/v12060670

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