PMID- 32661087
OWN - NLM
STAT- MEDLINE
DCOM- 20210618
LR  - 20210828
IS  - 1477-9137 (Electronic)
IS  - 0021-9533 (Print)
IS  - 0021-9533 (Linking)
VI  - 133
IP  - 16
DP  - 2020 Aug 27
TI  - Disc-associated proteins mediate the unusual hyperstability of the ventral disc 
      in Giardia lamblia.
LID - 10.1242/jcs.227355 [doi]
LID - jcs227355
AB  - Giardia lamblia, a widespread parasitic protozoan, attaches to the host 
      gastrointestinal epithelium by using the ventral disc, a complex microtubule (MT) 
      organelle. The 'cup-like' disc is formed by a spiral MT array that scaffolds 
      numerous disc-associated proteins (DAPs) and higher-order protein complexes. In 
      interphase, the disc is hyperstable and has limited MT dynamics; however, it 
      remains unclear how DAPs confer these properties. To investigate mechanisms of 
      hyperstability, we confirmed the disc-specific localization of over 50 new DAPs 
      identified by using both a disc proteome and an ongoing GFP localization screen. 
      DAPs localize to specific disc regions and many lack similarity to known 
      proteins. By screening 14 CRISPRi-mediated DAP knockdown (KD) strains for defects 
      in hyperstability and MT dynamics, we identified two strains - DAP5188KD and 
      DAP6751KD -with discs that dissociate following high-salt fractionation. Discs in 
      the DAP5188KD strain were also sensitive to treatment with the MT-polymerization 
      inhibitor nocodazole. Thus, we confirm here that at least two of the 87 known 
      DAPs confer hyperstable properties to the disc MTs, and we anticipate that other 
      DAPs contribute to disc MT stability, nucleation and assembly.
CI  - (c) 2020. Published by The Company of Biologists Ltd.
FAU - Nosala, Christopher
AU  - Nosala C
AUID- ORCID: 0000-0002-0179-8101
AD  - Department of Microbiology and Molecular Genetics, University of California, 
      Davis, One Shields Avenue, Davis, CA 95616, USA.
FAU - Hagen, Kari D
AU  - Hagen KD
AUID- ORCID: 0000-0001-9673-2481
AD  - Department of Microbiology and Molecular Genetics, University of California, 
      Davis, One Shields Avenue, Davis, CA 95616, USA.
FAU - Hilton, Nicholas
AU  - Hilton N
AD  - Department of Microbiology and Molecular Genetics, University of California, 
      Davis, One Shields Avenue, Davis, CA 95616, USA.
FAU - Chase, Tiffany M
AU  - Chase TM
AUID- ORCID: 0000-0001-9931-7697
AD  - Department of Microbiology and Molecular Genetics, University of California, 
      Davis, One Shields Avenue, Davis, CA 95616, USA.
FAU - Jones, Kelci
AU  - Jones K
AD  - Department of Microbiology and Molecular Genetics, University of California, 
      Davis, One Shields Avenue, Davis, CA 95616, USA.
FAU - Loudermilk, Rita
AU  - Loudermilk R
AUID- ORCID: 0000-0001-9970-5112
AD  - Department of Microbiology and Molecular Genetics, University of California, 
      Davis, One Shields Avenue, Davis, CA 95616, USA.
FAU - Nguyen, Kristofer
AU  - Nguyen K
AUID- ORCID: 0000-0001-8520-4835
AD  - Department of Microbiology and Molecular Genetics, University of California, 
      Davis, One Shields Avenue, Davis, CA 95616, USA.
FAU - Dawson, Scott C
AU  - Dawson SC
AUID- ORCID: 0000-0002-0843-1759
AD  - Department of Microbiology and Molecular Genetics, University of California, 
      Davis, One Shields Avenue, Davis, CA 95616, USA scdawson@ucdavis.edu.
LA  - eng
GR  - R01 AI077571/AI/NIAID NIH HHS/United States
GR  - T32 AI060555/AI/NIAID NIH HHS/United States
GR  - T32 GM007377/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20200827
PL  - England
TA  - J Cell Sci
JT  - Journal of cell science
JID - 0052457
RN  - 0 (Proteome)
RN  - 0 (Protozoan Proteins)
SB  - IM
MH  - *Giardia lamblia/genetics
MH  - Interphase
MH  - Microtubules
MH  - Organelles
MH  - Proteome
MH  - Protozoan Proteins/genetics
PMC - PMC7473642
OTO - NOTNLM
OT  - Giardia
OT  - MAP
OT  - MIP
OT  - Microtubule
OT  - Microtubule inner protein
OT  - Microtubule-associated protein
OT  - Organelle
COIS- Competing interestsThe authors declare no competing or financial interests.
EDAT- 2020/07/15 06:00
MHDA- 2021/06/22 06:00
PMCR- 2021/08/27
CRDT- 2020/07/15 06:00
PHST- 2018/10/31 00:00 [received]
PHST- 2020/06/29 00:00 [accepted]
PHST- 2020/07/15 06:00 [pubmed]
PHST- 2021/06/22 06:00 [medline]
PHST- 2020/07/15 06:00 [entrez]
PHST- 2021/08/27 00:00 [pmc-release]
AID - jcs.227355 [pii]
AID - JCS227355 [pii]
AID - 10.1242/jcs.227355 [doi]
PST - epublish
SO  - J Cell Sci. 2020 Aug 27;133(16):jcs227355. doi: 10.1242/jcs.227355.