Polypeptide 2A of human rhinovirus type 2: Identification as a protease and characterization by mutational analysis

Sommergruber, Wolfgang; Zorn, Manfred; Blaas, Dieter; Fessl, Friederike; Volkmann, Peter; Maurer‐Fogy, Ingrid; Pallai, Peter V.; Merluzzi, Vincent J.; Matteo, Martha; Skern, Tim; Kuechler, Ernst

doi:10.1016/0042-6822(89)90042-1

Cited by 86 publications

(74 citation statements)

References 30 publications

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“…The cleavage activity responsible for this scission has yet to be identified. The primary processing of the HAV polyprotein was found to resemble neither that of aphtho-and cardioviruses (Vakharia et al, 1987;Jackson, 1986), nor that of entero-and rhinoviruses Sommergruber et al, 1989). Therefore, it may be a novel type of polyprotein cleavage mediated not by an activity specific to the 2A region, but by 3C pr°.…”

Section: Discussionmentioning

confidence: 99%

Identification of hepatitis A virus non-structural protein 2B and its release by the major virus protease 3C

Gosert

Cassinotti

Siegl

et al. 1996

Journal of General Virology

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The RNA genome of hepatitis A virus (HAV) encodes a giant polyprotein that is putatively cleaved proteolytically into four structural and seven non-structural proteins. So far, most of the proposed non-structural proteins and their respective cleavage sites have not been identified. A vaccinia virus recombinant (vRGORF) containing the complete HAV ORF under the control of the bacteriophage T7 promoter was used to express HAY in recombinant animal cells (BT7-H) that constitutively expressed T7 DNA-dependent RNA polymerase. A HAV-specific 27.5 kDa expression product was identified as peptide 2B. The 27.5 kDa 2B antigen was also found in HAV-infected MRC-5 cells. The N-terminal amino acid residues of the new peptide 2B are Ala-Lys-Ile-Ser-Leu-Phe and polyprotein cleavage between 2A and 2B occurred at amino acids 836-837 (Gln-Ala). Furthermore, heterologous expression in the same system of regions P1-P2 and of the protease 3C (3C pr°) gene, showed that P1-P2 polyprotein is not cleaved autocatalytically but by 3C p'°. Hence, 3C vr° is effective in cleaving the polyprotein 2A-2B junction.

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Section: Discussionmentioning

confidence: 99%

Identification of hepatitis A virus non-structural protein 2B and its release by the major virus protease 3C

Gosert

Cassinotti

Siegl

et al. 1996

Journal of General Virology

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“…pro (35,38). Consequently, we replaced Gly or Ala at this position with Lys and compared the cleavage of these mutants with wildtype Nup62.…”

Section: Nup62 Is Cleaved At Multiple Sites In Cells Infected With Hrvmentioning

confidence: 99%

Specific Cleavage of the Nuclear Pore Complex Protein Nup62 by a Viral Protease

Park

Skern

Gustin

2010

Journal of Biological Chemistry

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“…lytic step is performed by the viral protein 2A, a protease that cleaves intramolecularly at its own amino terminus (Toyoda et al, 1986;Sommergruber et al, 1989). In addition to this initial cleavage of the nascent polyprotein, 2A proteases are also responsible for the cleavage of eIF4y (formerly p220), a subunit of the cap-binding complex eIF-4F.…”

mentioning

confidence: 99%

“…From a structural point of view, these viral proteases belong to the serine protease family, even though they utilize a cysteine as an active site nucleophile (Bazan et al, 1988(Bazan et al, , 1989Gorbalenya et al, 1989;Sommergruber et al, 1989;Hellen et al, 1991). The structural relationship with serine proteases was verified recently by the determination of the 3D structure of proReprint requests to: Tilman Voss, Boehringer-Ingelheim Research Vienna, Bender+Co, Dr. Boehringer-Gasse 5-11, A-1 121 Vienna, Austria; e-mail: voss@aimp.una.ac.at.…”

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confidence: 99%

Spectroscopic characterization of rhino viral protease 2a: Zn is essential for the structural integrity

1995

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Recently, protease 2A of human rhinovirus 2 (HRV2 2A) was shown to require a zinc ion for the formation of an active enzyme although zinc is not involved mechanistically. The data presented clearly show that the zinc ion bound to a picornaviral-specific motif represents an essential component of the native structure, probably representing a new Zn-binding motif. This structure, containing mostly &strand elements as shown by CD spectroscopy, changes drastically upon removal of zinc. The zinc-depleted form does represent an intermediate with mostly unchanged secondary structure, but not a fully denatured random coil as obtained by guanidinium hydrochloride. This is indicated by the blue-shifted fluorescence spectra and by CD. The native protein exhibited a cooperative phase transition at 53 "C. In contrast, the zinc-depleted form did not show any transition at all, again demonstrating the stabilizing role of the zinc ion. A structural intermediate was observed during thermal and pH denaturation that may represent a molten globule, as suggested by its ANS binding.

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Polypeptide 2A of human rhinovirus type 2: Identification as a protease and characterization by mutational analysis

Cited by 86 publications

References 30 publications

Identification of hepatitis A virus non-structural protein 2B and its release by the major virus protease 3C

Identification of hepatitis A virus non-structural protein 2B and its release by the major virus protease 3C

Specific Cleavage of the Nuclear Pore Complex Protein Nup62 by a Viral Protease

Spectroscopic characterization of rhino viral protease 2a: Zn is essential for the structural integrity

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