The rotavirus VP5*/VP8* conformational transition permeabilizes membranes to Ca 2 .
| العنوان: | The rotavirus VP5*/VP8* conformational transition permeabilizes membranes to Ca 2 . |
|---|---|
| المؤلفون: | De Sautu M; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 250 Longwood Avenue, Boston, MA 02115, USA.; Laboratory of Molecular Medicine, Boston Children's Hospital, Boston, MA 02115, USA., Herrmann T; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 250 Longwood Avenue, Boston, MA 02115, USA., Jenni S; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 250 Longwood Avenue, Boston, MA 02115, USA., Harrison SC; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 250 Longwood Avenue, Boston, MA 02115, USA.; Laboratory of Molecular Medicine, Boston Children's Hospital, Boston, MA 02115, USA.; Howard Hughes Medical Institute, Harvard Medical School, Boston, MA 02115, USA. |
| المصدر: | BioRxiv : the preprint server for biology [bioRxiv] 2023 Oct 16. Date of Electronic Publication: 2023 Oct 16. |
| نوع المنشور: | Preprint |
| اللغة: | English |
| بيانات الدورية: | Country of Publication: United States NLM ID: 101680187 Publication Model: Electronic Cited Medium: Internet NLM ISO Abbreviation: bioRxiv Subsets: PubMed not MEDLINE |
| مستخلص: | Rotaviruses infect cells by delivering into the cytosol a transcriptionally active inner capsid particle (a "double-layer particle": DLP). Delivery is the function of a third, outer layer, which drives uptake from the cell surface into small vesicles from which the DLPs escape. In published work, we followed stages of rhesus rotavirus (RRV) entry by live-cell imaging and correlated them with structures from cryogenic electron microscopy and tomography (cryo-EM and cryo-ET). The virus appears to wrap itself in membrane, leading to complete engulfment and loss of Ca 2+ from the vesicle produced by the wrapping. One of the outer-layer proteins, VP7, is a Ca 2+ -stabilized trimer; loss of Ca 2+ releases both outer-layer proteins from the particle. The other outer-layer protein, VP4, activated by cleavage into VP8* and VP5*, is a trimer that undergoes a large-scale conformational rearrangement, reminiscent of the transition that viral fusion proteins undergo to penetrate a membrane. The rearrangement of VP5* thrusts a 250-residue, C-terminal segment of each of the three subunits outward, while allowing the protein to remain attached to the virus particle and to the cell being infected. We proposed that this segment inserts into the membrane of the target cell, enabling Ca 2+ to cross. In the work reported here, we show the validity of key aspects of this proposed sequence. By cryo-EM studies of liposome-attached virions ("triple-layer particles": TLPs) and single-particle fluorescence imaging of liposome-attached TLPs, we confirm insertion of the VP4 C-terminal segment into the membrane and ensuing generation of a Ca 2+ "leak". The results allow us to formulate a molecular description of early events in entry. We also discuss our observations in the context of other work on double-strand RNA virus entry. |
| التعليقات: | Update in: PLoS Pathog. 2024 Apr 4;20(4):e1011750. (PMID: 38574119) |
| References: | J Virol. 1982 Dec;44(3):983-92. (PMID: 6294346) J Virol. 2001 Mar;75(5):2276-87. (PMID: 11160731) Proc Natl Acad Sci U S A. 2006 Oct 31;103(44):16496-501. (PMID: 17053074) J Virol. 2002 Oct;76(20):10512-7. (PMID: 12239329) J Mol Biol. 2001 Dec 14;314(5):985-92. (PMID: 11743716) Sci Adv. 2020 Apr 15;6(16):eaay6410. (PMID: 32494598) PLoS Pathog. 2014 Sep 11;10(9):e1004355. (PMID: 25211455) Nat Rev Microbiol. 2012 Jan 23;10(3):165-77. (PMID: 22266782) Nat Microbiol. 2021 Nov;6(11):1424-1432. (PMID: 34702979) Nature. 2021 Feb;590(7847):666-670. (PMID: 33442061) Elife. 2021 Jun 11;10:. (PMID: 34114559) Nature. 2012 Apr 15;485(7397):256-9. (PMID: 22504179) J Gen Virol. 1980 Jan;46(1):75-85. (PMID: 6243348) J Virol. 2017 Aug 24;91(18):. (PMID: 28701394) EMBO J. 2011 Jan 19;30(2):408-16. (PMID: 21157433) J Virol. 2014 Aug;88(16):9072-85. (PMID: 24899176) Nat Commun. 2019 May 17;10(1):2216. (PMID: 31101900) Elife. 2018 Mar 07;7:. (PMID: 29513216) J Virol. 2013 Jan;87(2):1115-22. (PMID: 23135722) J Struct Biol. 2016 Jan;193(1):1-12. (PMID: 26592709) J Struct Biol. 2015 Nov;192(2):216-21. (PMID: 26278980) J Struct Biol. 2017 Feb;197(2):102-113. (PMID: 27444392) J Virol. 2013 Jul;87(13):7255-64. (PMID: 23616650) J Mol Biol. 2019 Aug 9;431(17):3124-3138. (PMID: 31233764) Cell Host Microbe. 2023 Apr 12;31(4):604-615.e4. (PMID: 36996819) J Virol. 2010 Jun;84(12):6200-7. (PMID: 20375171) EMBO J. 1989 Jun;8(6):1695-703. (PMID: 2548854) Science. 2009 Jun 12;324(5933):1444-7. (PMID: 19520960) J Virol. 2022 Aug 24;96(16):e0062722. (PMID: 35924923) Nat Methods. 2012 Jun 28;9(7):676-82. (PMID: 22743772) J Virol. 1991 Sep;65(9):4720-7. (PMID: 1651404) J Virol. 2018 Nov 27;92(24):. (PMID: 30258012) Protein Sci. 2021 Jan;30(1):70-82. (PMID: 32881101) J Virol. 1983 Aug;47(2):267-75. (PMID: 6312065) J Virol. 2009 Jul;83(14):7004-14. (PMID: 19439475) Z Naturforsch C. 1973 Nov-Dec;28(11):693-703. (PMID: 4273690) J Struct Biol. 2005 Oct;152(1):36-51. (PMID: 16182563) Elife. 2014 Aug 13;3:e03665. (PMID: 25122622) Nat Methods. 2017 Apr;14(4):331-332. (PMID: 28250466) Nat Protoc. 2013 Jan;8(1):1-16. (PMID: 23222454) Nat Struct Mol Biol. 2016 Jan;23(1):74-80. (PMID: 26641711) J Virol. 2010 Feb;84(4):1764-70. (PMID: 20007281) J Struct Biol. 2012 Dec;180(3):519-30. (PMID: 23000701) |
| معلومات مُعتمدة: | R01 CA013202 United States CA NCI NIH HHS |
| تواريخ الأحداث: | Date Created: 20231031 Latest Revision: 20240416 |
| رمز التحديث: | 20240417 |
| مُعرف محوري في PubMed: | PMC10614792 |
| DOI: | 10.1101/2023.10.15.562449 |
| PMID: | 37905109 |
| قاعدة البيانات: | MEDLINE |
| DOI: | 10.1101/2023.10.15.562449 |
|---|