Comprehensive Analysis Reveals Dynamic and Evolutionary Plasticity of Rab GTPases and Membrane Traffic in
Cellular sophistication is not exclusive to multicellular organisms, and unicellular eukaryotes can resemble differentiated animal cells in their complex network of membrane-bound structures. These comparisons can be illuminated by genome-wide surveys of key gene families. We report a systematic analysis of Rabs in a complex unicellular Ciliate, including gene prediction and phylogenetic clustering, expression profiling based on public data, and Green Fluorescent Protein (GFP) tagging. Rabs are monomeric GTPases that regulate membrane traffic. Because Rabs act as compartment-specific determinants, the number of Rabs in an organism reflects intracellular complexity. The Tetrahymena Rab family is similar in size to that in humans and includes both expansions in conserved Rab clades as well as many divergent Rabs. Importantly, more than 90% of Rabs are expressed concurrently in growing cells, while only a small subset appears specialized for other conditions. By localizing most Rabs in living cells, we could assign the majority to specific compartments. These results validated most phylogenetic assignments, but also indicated that some sequence-conserved Rabs were co-opted for novel functions. Our survey uncovered a rare example of a nuclear Rab and substantiated the existence of a previously unrecognized core Rab clade in eukaryotes. Strikingly, several functionally conserved pathways or structures were found to be associated entirely with divergent Rabs. These pathways may have permitted rapid evolution of the associated Rabs or may have arisen independently in diverse lineages and then converged. Thus, characterizing entire gene families can provide insight into the evolutionary flexibility of fundamental cellular pathways.
Vyšlo v časopise:
Comprehensive Analysis Reveals Dynamic and Evolutionary Plasticity of Rab GTPases and Membrane Traffic in. PLoS Genet 6(10): e32767. doi:10.1371/journal.pgen.1001155
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1001155
Souhrn
Cellular sophistication is not exclusive to multicellular organisms, and unicellular eukaryotes can resemble differentiated animal cells in their complex network of membrane-bound structures. These comparisons can be illuminated by genome-wide surveys of key gene families. We report a systematic analysis of Rabs in a complex unicellular Ciliate, including gene prediction and phylogenetic clustering, expression profiling based on public data, and Green Fluorescent Protein (GFP) tagging. Rabs are monomeric GTPases that regulate membrane traffic. Because Rabs act as compartment-specific determinants, the number of Rabs in an organism reflects intracellular complexity. The Tetrahymena Rab family is similar in size to that in humans and includes both expansions in conserved Rab clades as well as many divergent Rabs. Importantly, more than 90% of Rabs are expressed concurrently in growing cells, while only a small subset appears specialized for other conditions. By localizing most Rabs in living cells, we could assign the majority to specific compartments. These results validated most phylogenetic assignments, but also indicated that some sequence-conserved Rabs were co-opted for novel functions. Our survey uncovered a rare example of a nuclear Rab and substantiated the existence of a previously unrecognized core Rab clade in eukaryotes. Strikingly, several functionally conserved pathways or structures were found to be associated entirely with divergent Rabs. These pathways may have permitted rapid evolution of the associated Rabs or may have arisen independently in diverse lineages and then converged. Thus, characterizing entire gene families can provide insight into the evolutionary flexibility of fundamental cellular pathways.
Zdroje
1. DerbyMC
GleesonPA
2007 New insights into membrane trafficking and protein sorting. Int Rev Cytol 261 47 116
2. MartensS
McMahonHT
2008 Mechanisms of membrane fusion: disparate players and common principles. Nat Rev Mol Cell Biol 9 543 556
3. DacksJB
FieldMC
2007 Evolution of the eukaryotic membrane-trafficking system: origin, tempo and mode. J Cell Sci 120 2977 2985
4. BonifacinoJS
GlickBS
2004 The mechanisms of vesicle budding and fusion. Cell 116 153 166
5. SegevN
2001 Ypt/rab gtpases: regulators of protein trafficking. Sci STKE 2001 RE11
6. SegevN
2001 Ypt and Rab GTPases: insight into functions through novel interactions. Curr Opin Cell Biol 13 500 511
7. MarkgrafDF
PeplowskaK
UngermannC
2007 Rab cascades and tethering factors in the endomembrane system. FEBS Lett 581 2125 2130
8. DacksJB
PedenAA
FieldMC
2009 Evolution of specificity in the eukaryotic endomembrane system. Int J Biochem Cell Biol 41 330 340
9. Pereira-LealJB
TeichmannSA
2005 Novel specificities emerge by stepwise duplication of functional modules. Genome Res 15 552 559
10. DhirV
GouldingD
FieldMC
2004 TbRAB1 and TbRAB2 mediate trafficking through the early secretory pathway of Trypanosoma brucei. Mol Biochem Parasitol 137 253 265
11. Pereira-LealJB
SeabraMC
2001 Evolution of the Rab family of small GTP-binding proteins. J Mol Biol 313 889 901
12. BockJB
MaternHT
PedenAA
SchellerRH
2001 A genomic perspective on membrane compartment organization. Nature 409 839 841
13. ZhangJ
SchulzeKL
HiesingerPR
SuyamaK
WangS
2007 Thirty-one flavors of Drosophila rab proteins. Genetics 176 1307 1322
14. LalK
FieldMC
CarltonJM
WarwickerJ
HirtRP
2005 Identification of a very large Rab GTPase family in the parasitic protozoan Trichomonas vaginalis. Mol Biochem Parasitol 143 226 235
15. Saito-NakanoY
LoftusBJ
HallN
NozakiT
2005 The diversity of Rab GTPases in Entamoeba histolytica. Exp Parasitol 110 244 252
16. AllenRD
FokAK
2000 Membrane trafficking and processing in Paramecium. Int Rev Cytol 198 277 318
17. AllenRD
SchroederCC
FokAK
1992 Endosomal system of Paramecium: coated pits to early endosomes. J Cell Sci 101 Pt 2 449 461
18. AllenRD
StaehelinLA
1981 Digestive system membranes: freeze-fracture evidence for differentiation and flow in Paramecium. J Cell Biol 89 9 20
19. SedarAW
PorterKR
1955 The fine structure of cortical components of Paramecium multimicronucleatum. J Biophys Biochem Cytol 1 583 604
20. ThompsonGAJr
NozawaY
1977 Tetrahymena: a system for studying dynamic membrane alterations within the eukaryotic cell. Biochim Biophys Acta 472 55 92
21. AllenRD
1978 Membranes of ciliates: ultrastructure, biochemistry and fusion. Membrane Fusion Amsterdam Elsevier/North-Holland Biomed. Press 657 763
22. FrankelJ
2000 Cell biology of Tetrahymena thermophila. Methods Cell Biol 6299432807 27 125
23. EisenJA
CoyneRS
WuM
WuD
ThiagarajanM
2006 Macronuclear genome sequence of the ciliate Tetrahymena thermophila, a model eukaryote. PLoS Biol 4 e286 doi:10.1371/journal.pbio.0040286
24. AuryJM
JaillonO
DuretL
NoelB
JubinC
2006 Global trends of whole-genome duplications revealed by the ciliate Paramecium tetraurelia. Nature 444 171 178
25. AdhiamboC
BlisnickT
ToutiraisG
DelannoyE
BastinP
2009 A novel function for the atypical small G protein Rab-like 5 in the assembly of the trypanosome flagellum. J Cell Sci 122 834 841
26. Pereira-LealJB
SeabraMC
2000 The mammalian Rab family of small GTPases: definition of family and subfamily sequence motifs suggests a mechanism for functional specificity in the Ras superfamily. J Mol Biol 301 1077 1087
27. MiaoW
XiongJ
BowenJ
WangW
LiuY
2009 Microarray analyses of gene expression during the Tetrahymena thermophila life cycle. PLoS ONE 4 e4429 doi:10.1371/journal.pone.0004429
28. StargellLA
KarrerKM
GorovskyMA
1990 Transcriptional regulation of gene expression in Tetrahymena thermophila. Nucl Acids Res 18 6637 6639
29. NelsenEM
DebaultLE
1978 Transformation in Tetrahymena pyriformis: description of an inducible phenotype. J Protozool 25 113 119
30. BannoY
SasakiN
NozawaY
1987 Secretion heterogeneity of lysosomal enzymes in Tetrahymena pyriformis. Exp Cell Res 170 259 268
31. MadingerCL
CollinsK
FieldsLG
TaronCH
BennerJS
2010 Constitutive secretion in Tetrahymena thermophila. Eukaryot Cell 9 674 681
32. RahamanA
EldeNC
TurkewitzAP
2008 A dynamin-related protein required for nuclear remodeling in Tetrahymena. Curr Biol 18 1227 1233
33. MartindaleDW
AllisCD
BrunsPJ
1982 Conjugation in Tetrahymena thermophila. A temporal analysis of cytological stages. Exp Cell Res 140 227 236
34. OriasJD
HamiltonEP
OriasE
1983 A microtubule meshwork associated with gametic pronucleus transfer across a cell-cell junction. Science 222 181 184
35. NelsenEM
WilliamsNE
YiH
KnaakJ
FrankelJ
1994 “Fenestrin” and conjugation in Tetrahymena thermophila. J Eukaryot Microbiol 41 483 495
36. CalzoneFJ
AngererRC
GorovskyMA
1983 Regulation of protein synthesis in Tetrahymena. Quantitative estimates of the parameters determining the rates of protein synthesis in growing, starved and starved-deciliated cells. J Biol Chem 258 6887 6898
37. JacobsME
DeSouzaLV
SamaranayakeH
PearlmanRE
SiuKW
2006 The Tetrahymena thermophila phagosome proteome. Eukaryot Cell 5 1990 2000
38. NilssonJR
1979 Phagotrophy in Tetrahymena.
LevandowskyM
HutnerSH
Biochemistry and Physiology of Protozoa New York Academic Press 339 379
39. AllenRD
FokAK
1980 Membrane recycling and endocytosis in Paramecium confirmed by horseradish peroxidase pulse-chase studies. J Cell Sci 45 131 145
40. NilssonJR
Van DeursB
1983 Coated pits and pinocytosis in Tetrahymena. J Cell Sci 63 209 222
41. AllenRD
1967 Fine structure, reconstruction and possible functions of components of the cortex of Tetrahymena pyriformis. J Protozool 14 553 565
42. EldeNC
MorganG
WineyM
SperlingL
TurkewitzAP
2005 Elucidation of Clathrin-Mediated Endocytosis in Tetrahymena Reveals an Evolutionarily Convergent Recruitment of Dynamin. PLoS Genet 1 e52 doi:10.1371/journal.pgen.0010052
43. KurzS
TiedtkeA
1993 The Golgi apparatus of Tetrahymena thermophila. J Eukaryot Microbiol 40 10 13
44. TurkewitzAP
2004 Out with a bang! Tetrahymena as a model system to study secretory granule biogenesis. Traffic 5 63 68
45. ZweifelE
SmithJ
RomeroD
GiddingsTHJr
WineyM
2009 Nested genes CDA12 and CDA13 encode proteins associated with membrane trafficking in the ciliate Tetrahymena thermophila. Eukaryot Cell 8 899 912
46. BowmanGR
EldeNC
MorganG
WineyM
TurkewitzAP
2005 Core Formation and the Acquisition of Fusion Competence are Linked During Secretory Granule Maturation in Tetrahymena. Traffic 6 303 323
47. CameronIL
BurtonAL
1969 On the cycle of the water expulsion vesicle in the ciliate Tetrahymena pyriformis. Trans Am Microsc Soc 88 386 393
48. DuF
EdwardsK
ShenZ
SunB
De LozanneA
2008 Regulation of contractile vacuole formation and activity in Dictyostelium. Embo J 27 2064 2076
49. TekleYI
GrantJR
KovnerAM
TownsendJP
KatzLA
2010 Identification of new molecular markers for assembling the eukaryotic tree of life. Mol Phylogenet Evol
50. NorianL
DragoiIA
O'HalloranT
1999 Molecular characterization of rabE, a developmentally regulated Dictyostelium homolog of mammalian rab GTPases. DNA Cell Biol 18 59 64
51. AllenRD
WolfRW
1979 Membrane recycling at the cytoproct of Tetrahymena. J Cell Sci 35 217 227
52. CetkovicH
MikocA
MullerWE
GamulinV
2007 Ras-like small GTPases form a large family of proteins in the marine sponge Suberites domuncula. J Mol Evol 64 332 341
53. CoyneRS
ThiagarajanM
JonesKM
WortmanJR
TallonLJ
2008 Refined annotation and assembly of the Tetrahymena thermophila genome sequence through EST analysis, comparative genomic hybridization, and targeted gap closure. BMC Genomics 9 562
54. ZufallRA
McGrathCL
MuseSV
KatzLA
2006 Genome architecture drives protein evolution in ciliates. Mol Biol Evol 23 1681 1687
55. RutherfordS
MooreI
2002 The Arabidopsis Rab GTPase family: another enigma variation. Curr Opin Plant Biol 5 518 528
56. KienleN
KloepperTH
FasshauerD
2009 Phylogeny of the SNARE vesicle fusion machinery yields insights into the conservation of the secretory pathway in fungi. BMC Evol Biol 9 19
57. KienleN
KloepperTH
FasshauerD
2009 Differences in the SNARE evolution of fungi and metazoa. Biochem Soc Trans 37 787 791
58. Pereira-LealJB
2008 The Ypt/Rab family and the evolution of trafficking in fungi. Traffic 9 27 38
59. EldeNC
LongM
TurkewitzAP
2007 A role for convergent evolution in the secretory life of cells. Trends Cell Biol 17 157 164
60. NachuryMV
LoktevAV
ZhangQ
WestlakeCJ
PeränenJ
2007 A core complex of BBS proteins cooperates with the GTPase Rab8 to promote ciliary membrane biogenesis. Cell 129 1201 1213
61. KissmehlR
SchildeC
WassmerT
DanzerC
NuehseK
2007 Molecular identification of 26 syntaxin genes and their assignment to the different trafficking pathways in Paramecium. Traffic 8 523 542
62. HorowitzS
GorovskyMA
1985 An unusual genetic code in nuclear genes of Tetrahymena. Proc Natl Acad Sci U S A 82 2452 2455
63. GuindonS
GascuelO
2003 A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol 52 696 704
64. HuelsenbeckJP
RonquistF
2001 MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics 17 754 755
65. RonquistF
HuelsenbeckJP
2003 MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19 1572 1574
66. FelsensteinJ
2006 Accuracy of coalescent likelihood estimates: do we need more sites, more sequences, or more loci? Mol Biol Evol 23 691 700
67. EdgarRC
2004 MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32 1792 1797
68. GouyM
GuindonS
GascuelO
2010 SeaView version 4: A multiplatform graphical user interface for sequence alignment and phylogenetic tree building. Mol Biol Evol 27 221 224
69. FelsensteinJ
1989 PHYLIP - Phylogeny Inference Package (Version 3.2). Cladistics 5 164 166
70. MaloneCD
FalkowskaKA
LiAY
GalantiSE
KanuruRC
2008 Nucleus-specific importin alpha proteins and nucleoporins regulate protein import and nuclear division in the binucleate Tetrahymena thermophila. Eukaryot Cell 7 1487 1499
71. ZachariasDA
ViolinJD
NewtonAC
TsienRY
2002 Partitioning of lipid-modified monomeric GFPs into membrane microdomains of live cells. Science 296 913 916
72. ShangY
SongX
BowenJ
CorstanjeR
GaoY
2002 A robust inducible-repressible promoter greatly facilitates gene knockouts, conditional expression, and overexpression of homologous and heterologous genes in Tetrahymena thermophila. Proc Natl Acad Sci U S A 99 3734 3739
73. GaertigJ
GorovskyMA
1992 Efficient mass transformation of Tetrahymena thermophila by electroporation of conjugants. Proc Natl Acad Sci USA 89 9196 9200
74. ChoiYY
JooMK
SohnYS
JeongB
2008 Significance of secondary structure in nanostructure formation and thermosensitivity of polypeptide block copolymers. Soft Matter 4 2383 2387
75. VidaTA
EmrSD
1995 A new vital stain for visualizing vacuolar membrane dynamics and endocytosis in yeast. J Cell Biol 128 779 792
76. StrackRL
StronginDE
BhattacharyyaD
TaoW
BermanA
2008 A noncytotoxic DsRed variant for whole-cell labeling. Nat Methods 5 955 957
77. ZhaoX
ClaudeA
ChunJ
ShieldsDJ
PresleyJF
2006 GBF1, a cis-Golgi and VTCs-localized ARF-GEF, is implicated in ER-to-Golgi protein traffic. J Cell Sci 119 3743 3753
78. BowmanGR
TurkewitzAP
2001 Analysis of a mutant exhibiting conditional sorting to dense core secretory granules in Tetrahymena thermophila. Genetics 159 1605 1616
79. HarrisE
YoshidaK
CardelliJ
BushJ
2001 Rab11-like GTPase associates with and regulates the structure and function of the contractile vacuole system in dictyostelium. J Cell Sci 114 3035 3045
80. WasmeierC
RomaoM
PlowrightL
BennettDC
RaposoG
2006 Rab38 and Rab32 control post-Golgi trafficking of melanogenic enzymes. J Cell Biol 175 271 281
81. AckersJP
DhirV
FieldMC
2005 A bioinformatic analysis of the RAB genes of Trypanosoma brucei. Mol Biochem Parasitol 141 89 97
82. QuevillonE
SpielmannT
BrahimiK
ChattopadhyayD
YeramianE
2003 The Plasmodium falciparum family of Rab GTPases. Gene 306 13 25
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