Effect of proteasome inhibitor clasto-lactacystin-beta-lactone on the proteome of the haloarchaeon Haloferax volcanii.
Proteasomes play key roles in a variety of eukaryotic cell functions, including translation, transcription, metabolism, DNA repair and cell-cycle control. The biological functions of these multicatalytic proteases in archaea, however, are poorly understood. In this study, Haloferax volcanii was used as a model to determine the influence the proteasome-specific inhibitor clasto-lactacystin-beta-lactone (cLbetaL) has on archaeal proteome composition. Addition of 20-30 microM cLbetaL had a widespread effect on the proteome, with a 38-42 % increase in the number of 2-D gel electrophoresis (2-DE) protein spots, from an average of 627 to 1036 spots. Protein identities for 17 of the spots that were easily separated by 2-DE and unique and/or increased 2- to 14-fold in the cLbetaL-treated cells were determined by tandem mass spectrometry (MS/MS). These included protein homologues of the DJ-1/ThiJ family, mobilization of sulfur system, translation elongation factor EF-1 A, ribosomal proteins, tubulin-like FtsZ, divalent metal ABC transporter, dihydroxyacetone kinase DhaL, aldehyde dehydrogenase and 2-oxoacid decarboxylase E1beta. Based on these results, inhibition of H. volcanii proteasomes had a global influence on proteome composition, including proteins involved in central functions of the cell.
Kirkland
P Aaron
PA
Department of Microbiology and Cell Science, University of Florida, Gainesville, 32611, USA.
Reuter
Christopher J
CJ
Department of Microbiology and Cell Science, University of Florida, Gainesville, 32611, USA.
Maupin-Furlow
Julie A
JA
Department of Microbiology and Cell Science, University of Florida, Gainesville, 32611, USA.
D001426:Bacterial Proteins; D019622:Haloferax volcanii / Q000737:chemistry* / Q000187:drug effects*; D007783:Lactones / Q000494:pharmacology*; D011480:Protease Inhibitors / Q000494:pharmacology*; D020543:Proteome
D001426:Bacterial Proteins; D007783:Lactones; D011480:Protease Inhibitors; D020543:Proteome; C098869:clasto-lactacystin beta-lactone