Viral proteins mimic host protein structure and function to redirect cellular processes and subvert innate defenses1. Small basic proteins compact and regulate both viral and cellular DNA genomes. Nucleosomes are the repeating units of cellular chromatin and play an important part in innate immune responses2. Viral-encoded core basic proteins compact viral genomes, but their impact on host chromatin structure and function remains unexplored. Adenoviruses encode a highly basic protein called protein VII that resembles cellular histones3. Although protein VII binds viral DNA and is incorporated with viral genomes into virus particles4,5, it is unknown whether protein VII affects cellular chromatin. Here we show that protein VII alters cellular chromatin, leading us to hypothesize that this has an impact on antiviral responses during adenovirus infection in human cells. We find that protein VII forms complexes with nucleosomes and limits DNA accessibility. We identified post-translational modifications on protein VII that are responsible for chromatin localization. Furthermore, proteomic analysis demonstrated that protein VII is sufficient to alter the protein composition of host chromatin. We found that protein VII is necessary and sufficient for retention in the chromatin of members of the high-mobility-group protein B family (HMGB1, HMGB2 and HMGB3). HMGB1 is actively released in response to inflammatory stimuli and functions as a danger signal to activate immune responses6,7. We showed that protein VII can directly bind HMGB1 in vitro and further demonstrated that protein VII expression in mouse lungs is sufficient to decrease inflammation-induced HMGB1 content and neutrophil recruitment in the bronchoalveolar lavage fluid. Together, our in vitro and in vivo results show that protein VII sequesters HMGB1 and can prevent its release. This study uncovers a viral strategy in which nucleosome binding is exploited to control extracellular immune signalling.

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We thank members of the Weitzman laboratory for insightful discussions and input, especially R. Dilley and B. Simpson for generating reagents. We also thank R. Panetierri and C. Koziol-White for providing precision-cut lung slices. We are grateful to D. Curiel for sharing recombinant protein-VII–GFP vectors and L. Gerace for anti-protein-VII antibodies. We thank the Penn Vector Core for assistance in purifying recombinant vectors, the Penn CDB Microscopy Core for imaging and FRAP assistance, and the CHOP Pathology core for immunostaining of mouse lungs. We thank members of the Black, Garcia and Worthen laboratories for technical help. We thank C. Bassing, I. Brodsky, J. Henao-Mejia, R. Kohli, C. Lopez, A. Resnick, S. Shin, K. Spindler and J. Weitzman for advice and critical reading of the manuscript. D.C.A. was supported in part by T32 CA115299 and F32 GM112414. N.J.P. was supported in part by T32 NS007180. N.S. was supported in part by funding from the American Cancer Society. Research was supported by grants from the National Institutes of Health (CA097093 to M.D.W., AI102577 and CA122677 to P.H., AI118891 and GM110174 to B.A.G., and GM082989 to B.E.B.), the Institute for Immunology of the University of Pennsylvania, and funds from the Children’s Hospital of Philadelphia (M.D.W.).

Author information

Author notes

    • Nikolina Sekulic

    Present address: Biotechnology Centre of Oslo and Department of Chemistry, University of Oslo, Oslo 0316, Norway.


  1. Department of Pathology and Laboratory Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania 19104, USA

    • Daphne C. Avgousti
    • , Katarzyna Kulej
    • , Emigdio D. Reyes
    •  & Matthew D. Weitzman
  2. Division of Cancer Pathobiology, Department of Pathology and Laboratory Medicine, Children’s Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, USA

    • Daphne C. Avgousti
    • , Christin Herrmann
    • , Katarzyna Kulej
    • , Neha J. Pancholi
    • , Joana Petrescu
    • , Emigdio D. Reyes
    •  & Matthew D. Weitzman
  3. Cell and Molecular Biology Graduate Group, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania 19104, USA

    • Christin Herrmann
    •  & Neha J. Pancholi
  4. Department of Biochemistry and Biophysics, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania 19104, USA

    • Nikolina Sekulic
    • , Ben E. Black
    •  & Benjamin A. Garcia
  5. Epigenetics Program, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania 19104, USA

    • Nikolina Sekulic
    • , Rosalynn C. Molden
    • , Ben E. Black
    •  & Benjamin A. Garcia
  6. Villanova University, Villanova, Pennsylvania 19085, USA

    • Joana Petrescu
  7. Division of Cell Pathology, Department of Pathology and Laboratory Medicine, Children’s Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, USA

    • Daniel Blumenthal
  8. Division of Pulmonary, Allergy, and Critical Care Medicine, Hospital of the University of Pennsylvania, and the Department of Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania 19104, USA

    • Andrew J. Paris
  9. Department of Molecular Genetics and Microbiology, School of Medicine, Stony Brook University, Stony Brook, New York 11794, USA

    • Philomena Ostapchuk
    •  & Patrick Hearing
  10. Protein and Proteomics Core, Children’s Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, USA

    • Steven H. Seeholzer
  11. Division of Neonatology, Children’s Hospital of Philadelphia, Philadelphia, and Department of Pediatrics, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania 19104, USA

    • G. Scott Worthen


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D.C.A. and M.D.W. conceived the project and designed experiments; D.C.A., C.H., N.S., J.P., N.J.P. and E.D.R. performed the experiments; D.C.A., C.H. and J.P. generated constructs and cell lines; K.K., R.C.M., S.H.S. and B.A.G. performed MS analysis; P.O. and P.H. generated Ad5-flox-VII virus and provided 293-Cre cell line; D.C.A. and D.B. performed the FRAP experiments; A.J.P. and G.S.W. conducted all mouse experiments; B.E.B. and B.A.G. designed experiments and interpreted the data; D.C.A. and M.D.W. interpreted the data and wrote the manuscript and all authors were involved in editing the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Matthew D. Weitzman.

All proteomic raw files have been deposited in the Chorus database under project number 1047 (https://chorusproject.org/).

Reviewer Information Nature thanks M. Bianchi and the other anonymous reviewer(s) for their contribution to the peer review of this work.

Extended data

Supplementary information

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  1. 1.

    Supplementary Figures

    This file contains the Source data gels for Figures 1d, e, f, 2a, 3b, c, j, k, 4b and Extended Data Figures 2a, b, c, d, e, f, g, 4a, b, c, 8a, g, 9c, f.

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  1. 1.

    Supplementary Table 1

    Summary of post-translational modifications found on histones upon expression of protein VII.

  2. 2.

    Supplementary Table 2

    Total list of proteins significantly changed upon protein VII expression identified by mass spectrometry analysis of high salt fractions. Table includes the log2 fold change of MaxQuant-derived iBAQ values obtained for protein VII-HA induced and uninduced highest salt fractions (600mM). Proteins with homoscedastic two-tailed t-test p-value smaller than 0.05 were considered as significantly altered between the two tested conditions. N/A defines not assigned t-test p-values; this is either due to the presence of the protein in only one condition, or if in one condition the protein was quantified in only one replicate. The number of peptides used for quantification was also highlighted. Commonly occurring contaminants, such as human keratins or trypsin, were removed from the final list.

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