Standing out in the field of IgY Immunotechnology

  • Home
    Home A full collection of all the Research Archive entries.
  • Years
    Years Sort entries by year.
  • Tags
    Tags Displays a list of tags that have been used in the blog.
  • Archives
    Archives Contains a list of research entries that were created previously.

RTA Occupancy of the Origin of Lytic Replication during Murine Gammaherpesvirus 68 Reactivation from B Cell Latency*

Posted by on in 2017
  • Font size: Larger Smaller
  • Hits: 758
  • Print

Santana AL1,2Oldenburg DG3Kirillov V4Malik L5Dong Q6Sinayev R7Marcu KB8,9,10,11White DW12Krug LT13Pathogens. 2017 Feb 16;6(1). pii: E9. doi: 10.3390/pathogens6010009.

1
The Ronald O. Perelman Department of Dermatology, New York University School of Medicine, New York, NY 10016, USA. Alexis.santana@nyumc.org.
2
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY 11794, USA. Alexis.santana@nyumc.org.
3
Gundersen Health System, La Crosse, WI 54601, USA. darby.oldenburg@gmail.com.
4
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY 11794, USA. varvara.kirillov@stonybrook.edu.
5
Department of Computer Science, Stony Brook University, Stony Brook, NY 11794, USA. laraib.malik@stonybrook.edu.
6
Program in Molecular and Cellular Biology, Stony Brook University, Stony Brook, NY 11794, USA. qiwen.dong@stonybrook.edu.
7
Applied Mathematics and Statistics, Stony Brook University, Stony Brook, NY 11794, USA. roman@rsinayev.com.
8
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY 11794, USA. kenneth.marcu@stonybrook.edu.
9
Biomedical Research Foundation Academy of Athens (BRFAA), Athens 115 27, Greece. kenneth.marcu@stonybrook.edu.
10
Biochemistry and Cell Biology Dept., Stony Brook University, Stony Brook, NY 11794, USA. kenneth.marcu@stonybrook.edu.
11
Department of Pathology, Health Sciences Center, Stony Brook University, Stony Brook, NY 11794, USA. kenneth.marcu@stonybrook.edu.
12
Gundersen Health System, La Crosse, WI 54601, USA. DWWhite@gundersenhealth.org.
13
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY 11794, USA. laurie.krug@stonybrook.edu.

Abstract

RTA, the viral Replication and Transcription Activator, is essential for rhadinovirus lytic gene expression upon de novo infection and reactivation from latency. Lipopolysaccharide (LPS)/toll-like receptor (TLR)4 engagement enhances rhadinovirus reactivation. We developed two new systems to examine the interaction of RTA with host NF-kappaB (NF-κB) signaling during murine gammaherpesvirus 68 (MHV68) infection: a latent B cell line (HE-RIT) inducible for RTA-Flag expression and virus reactivation; and a recombinant virus (MHV68-RTA-Bio) that enabled in vivo biotinylation of RTA in BirA transgenic mice. LPS acted as a second stimulus to drive virus reactivation from latency in the context of induced expression of RTA-Flag. ORF6, the gene encoding the single-stranded DNA binding protein, was one of many viral genes that were directly responsive to RTA induction; expression was further increased upon treatment with LPS. However, NF-κB sites in the promoter of ORF6 did not influence RTA transactivation in response to LPS in HE-RIT cells. We found no evidence for RTA occupancy of the minimal RTA-responsive region of the ORF6 promoter, yet RTA was found to complex with a portion of the right origin of lytic replication (oriLyt-R) that contains predicted RTA recognition elements. RTA occupancy of select regions of the MHV-68 genome was also evaluated in our novel in vivo RTA biotinylation system. Streptavidin isolation of RTA-Bio confirmed complex formation with oriLyt-R in LPS-treated primary splenocytes from BirA mice infected with MHV68 RTA-Bio. We demonstrate the utility of reactivation-inducible B cells coupled with in vivo RTA biotinylation for mechanistic investigations of the interplay of host signaling with RTA.

KEYWORDS:

NF-kappaB; gammaherpesvirus; latency; reactivation

PMID:
 
28212352
 
PMCID:
 
PMC5371897
 
DOI:
 
10.3390/pathogens6010009

*Buy them here:

Custom peptides, IgY antibodies (anti-Murine Gammaheresvirus-68 ORF59 (DNA polymerase processivity factor)) and Goat anti-Chicken IgY used in this publication were purchased from  Gallus Immunotech Inc. 

 

 

 

Last modified on