In Vivo Labelling of Adenovirus DNA Identifies Chromatin Anchoring and Biphasic Genome Replication

Journal of Virology
Tetsuro KomatsuHarald Wodrich

Abstract

Adenoviruses are DNA viruses with a lytic infection cycle. Following the fate of incoming as well as recently replicated genomes during infections is a challenge. In this study, we used the ANCHOR3 technology based on a bacterial partitioning system to establish a versatile in vivo imaging system for adenoviral genomes. The system allows the visualization of both individual incoming and newly replicated genomes in real time in living cells. We demonstrate that incoming adenoviral genomes are attached to condensed cellular chromatin during mitosis, facilitating the equal distribution of viral genomes in daughter cells after cell division. We show that the formation of replication centers occurs in conjunction with in vivo genome replication and determine replication rates. Visualization of adenoviral DNA revealed that adenoviruses exhibit two kinetically distinct phases of genome replication. Low-level replication occurred during early replication, while high-level replication was associated with late replication phases. The transition between these phases occurred concomitantly with morphological changes of viral replication compartments and with the appearance of virus-induced postreplication (ViPR) bodies, identified by the n...Continue Reading

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Citations

Nov 27, 2019·FEBS Letters·Kelsey L LynchDaphne C Avgousti
Nov 26, 2019·FEBS Letters·Matthew CharmanMatthew D Weitzman
Nov 24, 2019·FEBS Letters·Noémie Pied, Harald Wodrich
Feb 6, 2020·Viruses·Matthew Charman, Matthew D Weitzman
Mar 13, 2020·Journal of Virology·Irene Carlon-AndresHarald Wodrich
Feb 1, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Thorsten G MüllerBarbara Müller
Aug 25, 2020·Biochimica Et Biophysica Acta. Molecular Cell Research·Quentin NeversYves Gaudin
May 29, 2021·ACS Infectious Diseases·Sandrine Kappler-GratiasFranck Gallardo
May 23, 2021·Journal of Virological Methods·Charlotte Quentin-FroignantFranck Gallardo

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Methods Mentioned

BETA
fluorescence recovery after photobleaching
confocal microscopy
genetic modification
transfection
PCR
fluorescence microscopy
fluorescence-activated cell sorting

Software Mentioned

ANCHOR3
Addgene
PRISM7
OR3
ImageJ

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