PMID- 18931121
OWN - NLM
STAT- MEDLINE
DCOM- 20081208
LR  - 20181113
IS  - 1098-5530 (Electronic)
IS  - 0021-9193 (Print)
IS  - 0021-9193 (Linking)
VI  - 190
IP  - 24
DP  - 2008 Dec
TI  - Proteasomal components required for cell growth and stress responses in the 
      haloarchaeon Haloferax volcanii.
PG  - 8096-105
LID - 10.1128/JB.01180-08 [doi]
AB  - Little is known regarding the biological roles of archaeal proteases. The 
      haloarchaeon Haloferax volcanii is an ideal model for understanding these 
      enzymes, as it is one of few archaea with an established genetic system. In this 
      report, a series of H. volcanii mutant strains with markerless and/or conditional 
      knockouts in each known proteasome gene was systematically generated and 
      characterized. This included single and double knockouts of genes encoding the 
      20S core alpha1 (psmA), beta (psmB), and alpha2 (psmC) subunits as well as genes 
      (panA and panB) encoding proteasome-activating nucleotidase (PAN) proteins 
      closely related to the regulatory particle triple-A ATPases (Rpt) of eukaryotic 
      26S proteasomes. Our results demonstrate that 20S proteasomes are required for 
      growth. Although synthesis of 20S proteasomes containing either alpha1 or alpha2 
      could be separately abolished via gene knockout with little to no impact on 
      growth, conditional depletion of either beta alone or alpha1 and alpha2 together 
      rendered the cells inviable. In contrast, the PAN proteins were not essential 
      based on the robust growth of the panA panB double knockout strain. Deletion of 
      genes encoding either alpha1 or PanA did, however, render cells more sensitive to 
      growth on organic versus inorganic nitrogen sources and hypo-osmotic stress and 
      limited growth in the presence of l-canavanine. Abolishment of alpha1 synthesis 
      also had a severe impact on the ability of cells to withstand thermal stress. 
      This contrasted with what was seen for panA knockouts, which displayed enhanced 
      thermotolerance. Together, these results provide new and important insight into 
      the biological role of proteasomes in archaea.
FAU - Zhou, Guangyin
AU  - Zhou G
AD  - Department of Microbiology and Cell Science, University of Florida, Gainesville, 
      FL 32611-0700, USA.
FAU - Kowalczyk, David
AU  - Kowalczyk D
FAU - Humbard, Matthew A
AU  - Humbard MA
FAU - Rohatgi, Sunil
AU  - Rohatgi S
FAU - Maupin-Furlow, Julie A
AU  - Maupin-Furlow JA
LA  - eng
GR  - R01 GM057498/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20081017
PL  - United States
TA  - J Bacteriol
JT  - Journal of bacteriology
JID - 2985120R
RN  - 0 (Archaeal Proteins)
RN  - 0 (DNA, Archaeal)
RN  - 3HZV514J4B (Canavanine)
RN  - EC 3.4.25.1 (Proteasome Endopeptidase Complex)
RN  - EC 3.6.1.- (Adenosine Triphosphatases)
RN  - EC 3.6.4.- (PAN enzyme)
RN  - N762921K75 (Nitrogen)
SB  - IM
MH  - Adenosine Triphosphatases/genetics/*metabolism
MH  - Archaeal Proteins/genetics/*metabolism
MH  - Canavanine/metabolism
MH  - DNA, Archaeal/genetics
MH  - Gene Deletion
MH  - Gene Knockout Techniques
MH  - Haloferax volcanii/enzymology/genetics/*growth & development
MH  - Mutation
MH  - Nitrogen/metabolism
MH  - Phenotype
MH  - Promoter Regions, Genetic
MH  - Proteasome Endopeptidase Complex/genetics/*metabolism
MH  - Stress, Physiological
PMC - PMC2593203
EDAT- 2008/10/22 09:00
MHDA- 2008/12/17 09:00
PMCR- 2009/06/01
CRDT- 2008/10/22 09:00
PHST- 2008/10/22 09:00 [pubmed]
PHST- 2008/12/17 09:00 [medline]
PHST- 2008/10/22 09:00 [entrez]
PHST- 2009/06/01 00:00 [pmc-release]
AID - JB.01180-08 [pii]
AID - 1180-08 [pii]
AID - 10.1128/JB.01180-08 [doi]
PST - ppublish
SO  - J Bacteriol. 2008 Dec;190(24):8096-105. doi: 10.1128/JB.01180-08. Epub 2008 Oct 
      17.