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UV Radiation Induces Genome‐Mediated, Site‐Specific Cleavage in Viral Proteins
Author(s) -
Wigginton Krista Rule,
Menin Laure,
Sigstam Therese,
Gan Greg,
Cascella Michele,
Hamidane Hisham Ben,
Tsybin Yury O.,
Waridel Patrice,
Kohn Tamar
Publication year - 2012
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201100601
Subject(s) - cleavage (geology) , chemistry , capsid , cleavage factor , genome , bacteriophage , biophysics , rna , tandem mass spectrometry , mass spectrometry , biochemistry , biology , gene , escherichia coli , paleontology , chromatography , fracture (geology)
Much research has been dedicated to understanding the molecular basis of UV damage to biomolecules, yet many questions remain regarding the specific pathways involved. Here we describe a genome‐mediated mechanism that causes site‐specific virus protein cleavage upon UV irradiation. Bacteriophage MS2 was disinfected with 254 nm UV, and protein damage was characterized with ESI‐ and MALDI‐based FT‐ICR, Orbitrap, and TOF mass spectroscopy. Top‐down mass spectrometry of the products identified the backbone cleavage site as Cys46–Ser47 in the virus capsid protein, a location of viral genome–protein interaction. The presence of viral RNA was essential to inducing backbone cleavage. The similar bacteriophage GA did not exhibit site‐specific protein cleavage. Based on the major protein fragments identified by accurate mass analysis, a cleavage mechanism is proposed by radical formation. The mechanism involves initial oxidation of the Cys46 side chain followed by hydrogen atom abstraction from Ser47 C α . Computational protein QM/MM studies confirmed the initial steps of the radical mechanism. Collectively, this study describes a rare incidence of genome‐induced protein cleavage without the addition of sensitizers.