Intra- and inter-subunit disulfide bond formation is nonessential in adeno-associated viral capsids

Nagesh Pulicherla, Pradeep Kota, Nikolay V. Dokholyan, Aravind Asokan

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The capsid proteins of adeno-associated viruses (AAV) have five conserved cysteine residues. Structural analysis of AAV serotype 2 reveals that Cys289 and Cys361 are located adjacent to each other within each monomer, while Cys230 and Cys394 are located on opposite edges of each subunit and juxtaposed at the pentamer interface. The Cys482 residue is located at the base of a surface loop within the trimer region. Although plausible based on molecular dynamics simulations, intra- or inter-subunit disulfides have not been observed in structural studies. In the current study, we generated a panel of Cys-to-Ser mutants to interrogate the potential for disulfide bond formation in AAV capsids. The C289S, C361S and C482S mutants were similar to wild type AAV with regard to titer and transduction efficiency. However, AAV capsid protein subunits with C230S or C394S mutations were prone to proteasomal degradation within the host cells. Proteasomal inhibition partially blocked degradation of mutant capsid proteins, but failed to rescue infectious virions. While these results suggest that the Cys230/394 pair is critical, a C394V mutant was found viable, but not the corresponding C230V mutant. Although the exact nature of the structural contribution(s) of Cys230 and Cys394 residues to AAV capsid formation remains to be determined, these results support the notion that disulfide bond formation within the Cys289/361 or Cys230/394 pair appears to be nonessential. These studies represent an important step towards understanding the role of inter-subunit interactions that drive AAV capsid assembly.

Original languageEnglish (US)
Article numbere32163
JournalPloS one
Volume7
Issue number2
DOIs
StatePublished - Feb 28 2012

Fingerprint

Dependovirus
capsid
Capsid
disulfide bonds
Viruses
Disulfides
Capsid Proteins
mutants
coat proteins
Virus Assembly
Degradation
degradation
molecular dynamics
Protein Subunits
protein subunits
Mutant Proteins
Molecular Dynamics Simulation
virion
sulfides
Structural analysis

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

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title = "Intra- and inter-subunit disulfide bond formation is nonessential in adeno-associated viral capsids",
abstract = "The capsid proteins of adeno-associated viruses (AAV) have five conserved cysteine residues. Structural analysis of AAV serotype 2 reveals that Cys289 and Cys361 are located adjacent to each other within each monomer, while Cys230 and Cys394 are located on opposite edges of each subunit and juxtaposed at the pentamer interface. The Cys482 residue is located at the base of a surface loop within the trimer region. Although plausible based on molecular dynamics simulations, intra- or inter-subunit disulfides have not been observed in structural studies. In the current study, we generated a panel of Cys-to-Ser mutants to interrogate the potential for disulfide bond formation in AAV capsids. The C289S, C361S and C482S mutants were similar to wild type AAV with regard to titer and transduction efficiency. However, AAV capsid protein subunits with C230S or C394S mutations were prone to proteasomal degradation within the host cells. Proteasomal inhibition partially blocked degradation of mutant capsid proteins, but failed to rescue infectious virions. While these results suggest that the Cys230/394 pair is critical, a C394V mutant was found viable, but not the corresponding C230V mutant. Although the exact nature of the structural contribution(s) of Cys230 and Cys394 residues to AAV capsid formation remains to be determined, these results support the notion that disulfide bond formation within the Cys289/361 or Cys230/394 pair appears to be nonessential. These studies represent an important step towards understanding the role of inter-subunit interactions that drive AAV capsid assembly.",
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Intra- and inter-subunit disulfide bond formation is nonessential in adeno-associated viral capsids. / Pulicherla, Nagesh; Kota, Pradeep; Dokholyan, Nikolay V.; Asokan, Aravind.

In: PloS one, Vol. 7, No. 2, e32163, 28.02.2012.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Intra- and inter-subunit disulfide bond formation is nonessential in adeno-associated viral capsids

AU - Pulicherla, Nagesh

AU - Kota, Pradeep

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AU - Asokan, Aravind

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