Ultrastructure and function of viral replication complexes

Principal Investigator(s)

Dr. Montserrat Bárcena (CCB)Prof. Dr. Eric J. Snijder, Dr. Marjolein Kikkert

SARS-CoV RVNDescription of research

Viruses rely on the host cell’s infrastructure and metabolism during all stages of their replication cycle. It has become clear that the association of their RNA-synthesizing machinery with virus-induced membrane structures is a universal feature of mammalian +RNA viruses, and a potential target for broad-spectrum antiviral therapy. In infected cells, the viruses we study, such as the SARS-/MERS-coronaviruses and enteroviruses, induce replication organelles consisting of modified host membranes, including unique double-membrane structures that are derived from endoplasmic reticulum. Through the action of viral transmembrane proteins, corona- and enteroviruses hijack intracellular membranes to convert them into novel organelles supporting viral RNA synthesis. In addition, such replication organelles are thought to delay the detection of viral replication  by host cell sensors. Using a combination of biochemistry, structural and molecular biology, and advanced electron microscopy, we aim to dissect the formation, structure, and function of +RNA viral replication organelles. Knowledge derived from studying this early phase of infection will be essential for concepts aiming to block +RNA virus infection at an early stage.

This research project is carried out in close collaboration with the Section Electron Microscopy of the LUMC Department of Cell & Chemical Biology (CCB); Dr. Montserrat Bárcena, Prof. Bram J. Koster.

Research Topics

  • Structure and function of +RNA virus replication organelles
  • Advanced electron microscopy and tomography
  • Coronaviruses, arteriviruses and picornaviruses

Key Publications

  • Double-Membrane Vesicles as Platforms for Viral Replication. Wolff G, Melia CE, Snijder EJ, Bárcena M.Trends Microbiol. 2020 Jun 11:S0966-842X(20)30135-9. doi: 10.1016/j.tim.2020.05.009
  • A unifying structural and functional model of the coronavirus replication organelle: Tracking down RNA synthesis. Snijder EJ, Limpens RWAL, de Wilde AH, de Jong AWM, Zevenhoven-Dobbe JC, Maier HJ, Faas FFGA, Koster AJ, Bárcena M. PLoS Biol. 2020 Jun 8;18(6). doi: 10.1371/journal.pbio.3000715
  • A molecular pore spans the double membrane of the coronavirus replication organelle. Wolff G, Limpens RWAL, Zevenhoven-Dobbe JC, Laugks U, Zheng S, de Jong AWM, Koning RI, Agard DA, Grünewald K, Koster AJ, Snijder EJ, Bárcena M. Science. 2020 Aug 6. DOI: 10.1126/science.abd3629
  • The Origin, Dynamic Morphology, and PI4P-Independent Formation of Encephalomyocarditis Virus Replication Organelles. Melia CE, van der Schaar HM, de Jong AWM, Lyoo HR, Snijder EJ, Koster AJ, van Kuppeveld FJM, Bárcena M. MBio. 2018 Apr 17;9(2). pii: e00420-18. doi: 10.1128/mBio.00420-18.
  • Expression and Cleavage of Middle East Respiratory Syndrome Coronavirus nsp3-4 Polyprotein Induce the Formation of Double-Membrane Vesicles That Mimic Those Associated with Coronaviral RNA Replication. Oudshoorn D, Rijs K, Limpens RWAL, Groen K, Koster AJ, Snijder EJ, Kikkert M, Bárcena M. MBio. 2017 Nov 21;8(6). pii: e01658-17. doi: 10.1128/mBio.01658-17
  • Escaping Host Factor PI4KB Inhibition: Enterovirus Genomic RNA Replication in the Absence of Replication Organelles.Melia CE, van der Schaar HM, Lyoo H, Limpens RWAL, Feng Q, Wahedi M, Overheul GJ, van Rij RP, Snijder EJ, Koster AJ, Bárcena M, van Kuppeveld FJM. Cell Rep. 2017 Oct 17;21(3):587-599. doi: 10.1016/j.celrep.2017.09.068
  • Interaction of the innate immune system with positive-strand RNA virus replication organelles. Scutigliani EM, Kikkert M. Cytokine Growth Factor Rev. 2017 Oct;37:17-27
  • Antiviral Innate Immune Response Interferes with the Formation of Replication-Associated Membrane Structures Induced by a Positive-Strand RNA Virus. Oudshoorn D, van der Hoeven B, Limpens RW, Beugeling C, Snijder EJ, Bárcena M, Kikkert M. MBio. 2016 Dec 6;7(6).
  • Ultrastructural characterization of arterivirus replication structures: reshaping the endoplasmic reticulum to accommodate viral RNA synthesis.Knoops K, Bárcena M, Limpens RW, Koster AJ, Mommaas AM, Snijder EJ. J Virol. 2012; 86:2474-87.
  • The transformation of enterovirus replication structures: a three-dimensional study of single- and double-membrane compartments. Limpens RW, van der Schaar HM, Kumar D, Koster AJ, Snijder EJ, van Kuppeveld FJ, Bárcena M. mBio. 2011; 2:e00166-11.
  • SARS-coronavirus replication is supported by a reticulovesicular network of modified endoplasmic reticulum. Knoops K, Kikkert M, Worm SH, Zevenhoven-Dobbe JC, van der Meer Y, Koster AJ, Mommaas AM, Snijder EJ. PLoS Biol. 2008; 6:e226.
  • SARS-coronavirus replication/transcription complexes are membrane-protected and need a host factor for activity in vitro.van Hemert MJ, van den Worm SH, Knoops K, Mommaas AM, Gorbalenya AE, Snijder EJ. PLoS Pathog. 2008; 4:e1000054.