Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2002 Feb;8(2):188-201.
doi: 10.1017/s1355838202012785.

Determinants of the recognition of enteroviral cloverleaf RNA by coxsackievirus B3 proteinase 3C

Roland Zell  1 , Karim Sidigi, Enrico Bucci, Axel Stelzner, Matthias Görlach
Affiliations

Determinants of the recognition of enteroviral cloverleaf RNA by coxsackievirus B3 proteinase 3C

Roland Zell et al. RNA. 2002 Feb.

Abstract

The initiation of enteroviral positive-strand RNA synthesis requires the presence of a functional ribonucleoprotein complex containing a cloverleaf-like RNA secondary structure at the 5' end of the viral genome. Other components of the ribonucleoprotein complex are the viral 3CD proteinase (the precursor protein of the 3C proteinase and the 3D polymerase), the viral 3AB protein and the cellular poly(rC)-binding protein 2. For a molecular characterization of the RNA-binding properties of the enteroviral proteinase, the 3C proteinase of coxsackievirus B3 (CVB3) was bacterially expressed and purified. The recombinant protein is proteolytically active and forms a stable complex with in vitro-transcribed cloverleaf RNA of CVB3. The formation of stable complexes is also demonstrated with cloverleaf RNA of poliovirus (PV) 1, the first cloverleaf of bovine enterovirus (BEV) 1, and human rhinovirus (HRV) 2 but not with cloverleaf RNA of HRV14 and the second cloverleaf of BEV1. The apparent dissociation constants of the protein:RNA complexes range from approx. 1.7 to 4.6 microM. An electrophoretic mobility shift assay with subdomain D of the CVB3 cloverleaf demonstrates that this RNA is sufficient to bind the CVB3 3C proteinase. Binding assays using mutated versions of CVB3 and HRV14 cloverleaf RNAs suggest that the presence of structural features rather than a defined sequence motif of loop D are important for 3C proteinase-RNA interaction.

PubMed Disclaimer

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Cell. 1990 Oct 19;63(2):369-80 - PubMed
    1. Virology. 2001 Feb 1;280(1):80-6 - PubMed
    1. Virus Res. 1992 Jul;24(2):187-96 - PubMed
    1. Protein Eng. 1992 Apr;5(3):191-5 - PubMed
    1. J Biol Chem. 1993 Dec 5;268(34):25735-9 - PubMed

Publication types

MeSH terms

LinkOut - more resources