Reversion to wildtype of a mutated and nonfunctional coxsackievirus B3CRE(2C).

Item request has been placed! ×
Item request cannot be made. ×
loading   Processing Request
Read More Add to Saved list
  • Author(s): Smithee S;Smithee S;Smithee S; Tracy S; Tracy S; Chapman NM; Chapman NM
  • Source:
    Virus research [Virus Res] 2016 Jul 15; Vol. 220, pp. 136-49. Date of Electronic Publication: 2016 Apr 26.
  • Publication Type:
    Journal Article; Research Support, Non-U.S. Gov't; Research Support, N.I.H., Extramural
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: Elsevier Science Country of Publication: Netherlands NLM ID: 8410979 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1872-7492 (Electronic) Linking ISSN: 01681702 NLM ISO Abbreviation: Virus Res Subsets: MEDLINE
    • Publication Information:
      Original Publication: Amsterdam : Elsevier Science, c1984-
    • Subject Terms:
      Base Sequence* ; Mutation* ; Sequence Deletion*; Carrier Proteins/*genetics ; Enterovirus B, Human/*genetics ; Viral Nonstructural Proteins/*genetics; Animals ; Carrier Proteins/metabolism ; Coxsackievirus Infections/virology ; Enterovirus B, Human/metabolism ; Gene Expression ; HeLa Cells ; Heart/virology ; Humans ; Male ; Mice ; RNA, Viral/genetics ; Spleen/virology ; Transcription, Genetic ; Viral Nonstructural Proteins/metabolism ; Virus Replication
    • Abstract:
      The cis-acting replication element (CRE) in the 2C protein coding region [CRE(2C)] of enteroviruses (EV) facilitates the addition of two uridine residues (uridylylation) onto the virus-encoded protein VPg in order for it to serve as the RNA replication primer. We demonstrated that coxsackievirus B3 (CVB3) is replication competent in the absence of a native (uridylylating) CRE(2C) and also demonstrated that lack of a functional CRE(2C) led to generation of 5' terminal genomic deletions in the CVB3 CRE-knock-out (CVB3-CKO) population. We asked whether reversion of the mutated CRE(2C) occurred, thus permitting sustained replication, and when were 5' terminal deletions generated during replication. Virions were isolated from HeLa cells previously electroporated with infectious CVB3-CKO T7 transcribed RNA or from hearts and spleens of mice after transfection with CVB3-CKO RNA. Viral RNA was isolated in order to amplify the CRE(2C) coding region and the genomic 5' terminal sequences. Sequence analysis revealed reversion of the CVB3-CKO sequence to wildtype occurs by 8 days post-electroporation of HeLa cells and by 20days post-transfection in mice. However, 5' terminal deletions evolve prior to these times. Reversion of the CRE(2C) mutations to wildtype despite loss of the genomic 5' termini is consistent with the hypothesis that an intact CRE(2C) is inherently vital to EV replication even when it is not enabling efficient positive strand initiation.
      (Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.)
    • Contributed Indexing:
      Keywords: CRE(2C); Coxsackievirus; Enterovirus; Reversion; Terminal deletion
    • Accession Number:
      0 (Carrier Proteins)
      0 (RNA, Viral)
      0 (Viral Nonstructural Proteins)
      EC 3.6.1.- (2C protein, viral)
    • Publication Date:
      Date Created: 20160501 Date Completed: 20171206 Latest Revision: 20171218
    • Publication Date:
      20240829
    • Accession Number:
      10.1016/j.virusres.2016.04.016
    • Accession Number:
      27130630