title: Proteomics of secretory and endocytic organelles in Giardia lamblia.
abstract: Giardia lamblia is a flagellated protozoan enteroparasite transmitted as an environmentally resistant cyst. Trophozoites attach to the small intestine of vertebrate hosts and proliferate by binary fission. They access nutrients directly via uptake of bulk fluid phase material into specialized endocytic organelles termed peripheral vesicles (PVs), mainly on the exposed dorsal side. When trophozoites reach the G2/M restriction point in the cell cycle they can begin another round of cell division or encyst if they encounter specific environmental cues. They induce neogenesis of Golgi-like organelles, encystation-specific vesicles (ESVs), for regulated secretion of cyst wall material. PVs and ESVs are highly simplified and thus evolutionary diverged endocytic and exocytic organelle systems with key roles in proliferation and transmission to a new host, respectively. Both organelle systems physically and functionally intersect at the endoplasmic reticulum (ER) which has catabolic as well as anabolic functions. However, the unusually high degree of sequence divergence in Giardia rapidly exhausts phylogenomic strategies to identify and characterize the molecular underpinnings of these streamlined organelles. To define the first proteome of ESVs and PVs we used a novel strategy combining flow cytometry-based organelle sorting with in silico filtration of mass spectrometry data. From the limited size datasets we retrieved many hypothetical but also known organelle-specific factors. In contrast to PVs, ESVs appear to maintain a strong physical and functional link to the ER including recruitment of ribosomes to organelle membranes. Overall the data provide further evidence for the formation of a cyst extracellular matrix with minimal complexity. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium with the dataset identifier PXD000694.
authors.affiliation: Institute of Parasitology, University of Zurich, Zurich, Switzerland.
authors.forename: Petra B
authors.initials: PB
authors.lastname: Wampfler
authors.affiliation: Institute of Experimental Immunology, University of Zurich, Zurich, Switzerland.
authors.forename: Vinko
authors.initials: V
authors.lastname: Tosevski
authors.affiliation: Functional Genomics Center Zurich, Zurich, Switzerland.
authors.forename: Paolo
authors.initials: P
authors.lastname: Nanni
authors.affiliation: Institute of Parasitology, University of Zurich, Zurich, Switzerland; Institute of Parasitology, University of Bern, Bern, Switzerland.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Spycher
authors.affiliation: Institute of Parasitology, University of Zurich, Zurich, Switzerland.
authors.forename: Adrian B
authors.initials: AB
authors.lastname: Hehl
mesh_terms: D016000:Cluster Analysis; D030562:Databases, Protein; D004705:Endocytosis; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000378:metabolism; D016829:Giardia lamblia / Q000378:metabolism* / Q000648:ultrastructure; D013058:Mass Spectrometry; D058977:Molecular Sequence Annotation; D014176:Protein Biosynthesis; D040901:Proteomics; D015800:Protozoan Proteins / Q000378:metabolism*; D015203:Reproducibility of Results; D012270:Ribosomes / Q000378:metabolism / Q000648:ultrastructure; D022142:Secretory Vesicles / Q000378:metabolism* / Q000648:ultrastructure; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes; D015800:Protozoan Proteins
references.citation: J Eukaryot Microbiol. 2007 Mar-Apr;54(2):154-60
references.pmid: 17403156
references.citation: Eukaryot Cell. 2009 Nov;8(11):1665-76
references.pmid: 19749174
references.citation: Nature. 2003 Nov 13;426(6963):172-6
references.pmid: 14614504
references.citation: J Cell Sci. 2009 Aug 15;122(Pt 16):2846-56
references.pmid: 19622633
references.citation: Chem Biol Interact. 2003 Feb 1;143-144:247-53
references.pmid: 12604210
references.citation: J Cell Biol. 2008 Jun 30;181(7):1095-105
references.pmid: 18573918
references.citation: J Biol Chem. 2003 Jun 13;278(24):21701-8
references.pmid: 12686559
references.citation: PLoS One. 2006 Dec 20;1:e44
references.pmid: 17183673
references.citation: PLoS One. 2013 Dec 31;8(12):e83207
references.pmid: 24391747
references.citation: Mol Biol Cell. 2003 Apr;14(4):1433-47
references.pmid: 12686599
references.citation: Traffic. 2008 Sep;9(10):1698-716
references.pmid: 18637903
references.citation: Mol Biochem Parasitol. 1992 Dec;56(2):301-9
references.pmid: 1484552
references.citation: Gen Comp Endocrinol. 2012 Jan 1;175(1):1-9
references.pmid: 21907200
references.citation: Proteomics. 2010 Nov;10(22):3935-56
references.pmid: 21080488
references.citation: Int J Parasitol. 2011 Apr;41(5):471-80
references.pmid: 21296082
references.citation: Mol Biochem Parasitol. 2008 May;159(1):44-53
references.pmid: 18346800
references.citation: J Biol Chem. 1995 Dec 8;270(49):29307-13
references.pmid: 7493963
references.citation: Mol Cell Proteomics. 2008 Sep;7(9):1651-67
references.pmid: 18504258
references.citation: Nucleic Acids Res. 2013 Jan;41(Database issue):D1063-9
references.pmid: 23203882
references.citation: Mol Microbiol. 2011 Mar;79(5):1204-19
references.pmid: 21205007
references.citation: J Biol Chem. 2003 Jul 4;278(27):24837-48
references.pmid: 12711599
references.citation: PLoS Negl Trop Dis. 2010 Jun 01;4(6):e697
references.pmid: 20532229
references.citation: J Biol Chem. 2008 Dec 19;283(51):35996-6010
references.pmid: 18930915
references.citation: Clin Microbiol Rev. 2001 Jul;14(3):447-75
references.pmid: 11432808
references.citation: Mol Biochem Parasitol. 2011 Feb;175(2):169-80
references.pmid: 21075147
references.citation: Front Biosci (Elite Ed). 2012 Jan 01;4:1898-909
references.pmid: 22202006
references.citation: Nat Protoc. 2009;4(1):44-57
references.pmid: 19131956
references.citation: Biotechniques. 2005 Dec;39(6):809-10, 812
references.pmid: 16382896
references.citation: Science. 2007 Sep 28;317(5846):1921-6
references.pmid: 17901334
references.citation: Eur J Cell Biol. 1990 Oct;53(1):142-53
references.pmid: 2076701
references.citation: Parasitol Int. 2008 Jun;57(2):97-104
references.pmid: 18180199
references.citation: PLoS Negl Trop Dis. 2010 May 11;4(5):e677
references.pmid: 20485485
references.citation: Cell. 1997 May 2;89(3):437-44
references.pmid: 9150143
references.citation: Mol Biol Cell. 2004 Jul;15(7):3053-60
references.pmid: 15107467
references.citation: Mol Microbiol. 2004 Jul;53(1):19-28
references.pmid: 15225300
references.citation: Exp Parasitol. 1989 Feb;68(2):238-41
references.pmid: 2647510
references.citation: Traffic. 2004 Feb;5(2):63-8
references.pmid: 14690495
references.citation: Nature. 2008 Dec 11;456(7223):750-4
references.pmid: 19079052
references.citation: Mol Biochem Parasitol. 1989 Jan 15;32(2-3):121-31
references.pmid: 2927442
references.citation: Nat Commun. 2013;4:2493
references.pmid: 24042146
references.citation: J Biol Chem. 2004 May 7;279(19):20242-9
references.pmid: 14996840
references.citation: Mass Spectrom Rev. 2009 Sep-Oct;28(5):844-67
references.pmid: 19301366
references.citation: Eukaryot Cell. 2010 Oct;9(10):1566-76
references.pmid: 20693303
references.citation: Biochem J. 2010 Apr 28;428(1):33-45
references.pmid: 20199400
references.citation: Exp Parasitol. 1994 Nov;79(3):236-49
references.pmid: 7525336
references.citation: Mol Biochem Parasitol. 2010 Nov;174(1):62-5
references.pmid: 20570699
references.citation: J Biol Chem. 2006 Mar 17;281(11):7595-604
references.pmid: 16407213
references.citation: J Biol Chem. 2003 Feb 21;278(8):6420-6
references.pmid: 12466276
references.citation: Mol Biochem Parasitol. 2004 Aug;136(2):173-80
references.pmid: 15478797
references.citation: J Struct Biol. 1998 Nov;123(3):225-35
references.pmid: 9878577
references.citation: Cell Microbiol. 2013 Apr;15(4):537-53
references.pmid: 23094658
references.citation: Cell. 2001 Aug 24;106(4):499-509
references.pmid: 11525735
references.citation: J Cell Sci. 1996 Jul;109 ( Pt 7):1909-17
references.pmid: 8832413
references.citation: J Biol Chem. 2002 Mar 8;277(10):8474-81
references.pmid: 11773053
references.citation: Mol Microbiol. 1995 Mar;15(5):955-63
references.pmid: 7596296
references.citation: Int J Parasitol. 2008 Sep;38(11):1297-304
references.pmid: 18625508
references.citation: Trends Cell Biol. 2014 Jan;24(1):35-43
references.pmid: 24119662
references.citation: Proteomics. 2008 Sep;8(18):3848-61
references.pmid: 18704904
references.citation: PLoS Pathog. 2010 Apr 08;6(4):e1000835
references.pmid: 20386711
references.citation: Anal Biochem. 1999 Feb 15;267(2):344-50
references.pmid: 10036140
references.citation: Traffic. 2008 Jan;9(1):57-71
references.pmid: 17892527
references.citation: Biochem Biophys Res Commun. 2004 Nov 19;324(3):1069-80
references.pmid: 15485664
references.citation: Mol Microbiol. 1998 Dec;30(5):979-89
references.pmid: 9988475
references.citation: Mol Biol Cell. 2000 May;11(5):1789-800
references.pmid: 10793152
references.citation: J Protozool. 1968 Feb;15(1):26-30
references.pmid: 5643476
references.citation: J Struct Biol. 2003 Aug;143(2):153-63
references.pmid: 12972352
references.citation: Proc Natl Acad Sci U S A. 2005 Feb 1;102(5):1548-53
references.pmid: 15665075
references.citation: Exp Parasitol. 1988 Feb;65(1):141-7
references.pmid: 3276550
references.citation: PLoS Pathog. 2010 Aug 19;6(8):e1001059
references.pmid: 20808847
references.citation: PLoS One. 2011 Feb 24;6(2):e17285
references.pmid: 21390322
references.citation: EMBO J. 1993 Feb;12(2):749-58
references.pmid: 8440262
references.citation: Glycobiology. 2002 Aug;12(8):499-505
references.pmid: 12145190
references.citation: J Biol Chem. 2008 Jun 27;283(26):18024-31
references.pmid: 18445589
references.citation: FEMS Microbiol Lett. 2000 Aug 15;189(2):271-3
references.pmid: 10930750
references.citation: J Protozool. 1987 Feb;34(1):80-3
references.pmid: 3572844