The Golgi Apparatus: State of the art 110 years after Camillo Golgi’s discovery

713 Pages·2008·7.94 MB·English

by  Alexander A. Mironov

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Alexander A.MironovandMargit Pavelka(eds.)
The Golgi Apparatus
State of the art 110 years after Camillo Golgi’s discovery
SpringerWienNewYork
Prof. Dr. Alexander A. Mironov
ConsorzioMarioNegriSud
LaboratoryofIntracellularTraffic
DepartmentofCellBiologyandOncology,
S.MariaImbaro(Chieti),Italy
Prof. Dr. Margit Pavelka
DepartmentofCellBiologyandUltrastructureResearch
InstituteofHistologyandEmbryology
CenterforAnatomyandCellBiology
MedicalUniversityofVienna
Vienna,Austria
GedrucktmitUnterstu€tzungdesBundesministeriumsfu€rWissenschaftundForschung
inWien
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Preface * V
Preface
In 2008, we celebrate the 110th anniversary of the first description of the
complexapparatoreticolareinternobyCamilloGolgiintheBollettinodella
Societa(cid:2) Medico-ChirurgicadiPavia.ThebiographyofCamilloGolgi,andthe
GolgiapparatushistoryhaveimpressivelybeendescribedbyPaoloMazzarello
in his book “The Hidden Structure. A Scientific Biography of Camillo Golgi”
(OxfordUniversityPress,1999).Duringthe20thcentury,CamilloGolgi’sdiscov-
ery hadachangingupanddownanduphistory,timelybeingassessedasan
artefact,andthenagaincomingintothecentrestageofcellbiologicresearch.
Today,itiswellestablishedthattheGolgiapparatusconstitutesamaincross-
roadsintheintracellulartransportroutesofthebiosynthetic,endocytic,and
recyclingsystems.Duringthepastdecades,multiplenewdiscoveriescontribut-
edtotheunderstandingoftheorganizationandthefunctionsofthecomplex
organelle(seeChapter1.1).In1997,theexcellentbookabouttheGolgiap-
paratuseditedbyJ.RothandE.Bergerprovidedacomprehensivesummariz-
ingpresentationofthestateofresearch100yearsafterthefirstdescriptionof
theorganelle.Now,afterfurther10years,itisnecessarytosummarizeagain
whatitisknownaboutthecomplexorganizationofCamilloGolgi’sapparatus.
Ourbookisanattempttobringtogethermultiplenewresultsobtainedby
differenttechniques,andaddressingdifferentaspectsoftheGolgiapparatus
andintracellulartransport.Wearehopefulthatthepresentationofthestate
oftheart110yearsafterCamilloGolgi’sdiscoveryofthecomplexapparato
reticolareinternowillleadtoanimprovedunderstanding,novelinsights,and
newperspectivesforfutureresearch.
Acknowledgements
The editors cordially thank all authors of the Chapters, all our colleagues
involvedintheworkspresentedintheChaptersofthisbookwrittenbyusand
byothers.WethanktheSpringerCompanyforthepossibilitytopublishthis
book and in particular we thank Mag. Franziska Brugger, Mag. Angelika
Heller,andMrs.UrsulaSzorgerforahugehelpinourwork.A.M.isespecially
thankfultoDr.A.FusellaandD.Gaindomenicoforthetechnicalhelpandto
ChrisBerrieandRamanParashuramanforcriticalreadingofthemanuscripts,
inwhichA.M.isaco-author.
A.A.MironovandM.Pavelka
S.MariaImbaro/Vienna,May2008
Moviescanbeviewedonlineat:
www.springer.com/springerwiennewyork/lifeþsciences/book/978-3-211-76309-4
Contents * VII
Contents
Introduction ............................................... 1
1. Generalconsiderations .................................... 5
1.1. TheGolgiapparatusandmaindiscoveries inthefieldof
intracellulartransport................................ 7
1.2. TheGolgiapparatusasacrossroadsinintracellulartraffic... 16
2. MainmachineriesoperatingattheGolgiapparatus ............. 41
2.1. SNAREs............................................ 43
2.2. Rabs.............................................. 66
2.3. COPII ............................................. 78
2.4. COPI:mechanismsandtransportroles................... 87
2.5. ArfsandArls:modelsforArffamilymembersin
membranetrafficattheGolgi .........................106
2.6. COGcomplex.......................................120
2.7. TheTRAPPcomplex..................................130
2.8. TheroleofCa2þintheregulationofintracellular
transport ..........................................143
2.9. Golgiglycosylationenzymes...........................161
2.10. NucleotidesugartransportersoftheGolgiapparatus.......190
2.11. Luminallectins......................................207
2.12. TheGolgiribbonandthefunctionofthegolgins..........223
2.13. Functionalcrosstalkbetweenmembranetrafficking
andcellsignalling...................................247
2.14. Theroleofthecytoskeletoninthestructureandfunction
oftheGolgiapparatus ...............................270
2.15. Thedynamin–cortactincomplexasamediatorofvesicle
formationatthetrans-Golginetwork ...................301
2.16. Thegeometryoforganellesofthesecretorypathway ......314
3. Maintransportsteps ......................................331
3.1. ER-to-Golgitransport ................................333
3.2. Intra-Golgitransport.................................342
3.3. Structureanddomainorganizationofthe
trans-Golginetwork .................................358
3.4. Golgi-to-PMtransport................................375
3.5. Proteintransportfromthetrans-Golginetwork
toendosomes ......................................388
VIII * Contents
3.6. Thetransportofsolublelysosomalhydrolases fromthe
Golgicomplextolysosomes ...........................402
3.7. Transportoflysosomalmembraneproteins from
theGolgicomplextolysosomes........................414
3.8. Retrogradeendosome-to-TGNtransport .................425
3.9. Retrogradeplasmamembrane-to-Golgiapparatus
transport ..........................................459
3.10. Interactionsbetweenendocytosis andsecretorytransport ..475
3.11. Originsoftheregulatedsecretorypathway ..............485
3.12. Secretionandendocytosisinendothelialcells.............520
3.13. Formationofmucingranules..........................535
3.14. Golgiapparatusandepithelialcellpolarity...............563
3.15. Golgiapparatusinheritance...........................580
4. Peculiaritiesofintracellulartransportindifferentorganisms ......609
4.1. FeaturesoftheplantGolgiapparatus...................611
4.2. YeastGolgiapparatus................................623
4.3. MorphodynamicsoftheyeastGolgiapparatus............630
4.4. StructureandfunctionoftheGolgiorganelle
inparasiticprotists ..................................647
4.5. EvolutionoftheGolgicomplex ........................675
5. Generalconclusions.......................................693
Contributorsinalphabeticalorder..............................697
Index.....................................................706
Introduction * 1
Introduction
AlexanderA.MironovandMargitPavelka
Allcellssecreteadiversityofmacromoleculestomodifytheirenvironmentor
toprotectthemselves.Ontheotherhand,thereisthenecessitytoreplace
membrane proteins and lipids that are being constantly degraded in com-
partments of the secretory and endocytic pathways. Therefore, eukaryotic
cells synthesize proteins for either export (secretion) or delivery to the
secretory and endocytic compartments and to the plasma membrane (PM)
forreplacementofdegradedproteinsandlipids.Thesynthesisiscarriedout
byribosomesattachedtothecytosolicsurfaceoftheendoplasmicreticulum
(ER).Thesynthesisofmostofcellularlipidsandallfattyacidsalsooccursinthe
smoothER.Duringorafterthesynthesis,thepolypeptidechainscontaining
transmembranedomainscomposedofhydrophobicaminoacidsareinserted
intotheERmembrane,whereassolubleproteinsaretransferredintotheER
lumen.Afterthecleavageoftheirleadinghydrophobicsignalpeptidesand
afterproteinfoldingboththegroupsofproteinsaretransportedalongthe
secretorypathway.
TheGolgiapparatus(GA)isthecentralstationalongthispathway.While
passingthrough theGA,proteinsandlipids undergo posttranslational mod-
ifications (mainly glycosylation by Golgi glycosidases and transferases) and
sorting.ThetransportofproteinsandlipidsfromtheERtotheirdestinations
maybecarriedoutinseveralways:viathedissociationmechanism,thepro-
gressionmechanism,and/orthelateraldiffusionmechanism(seeChapter1.2).
Overthepast30years,thefieldofintracellulartraffichasseentremendous
advancestowardstheidentificationoftherelevantmolecularmachineries.
Substantialprogressintheisolation,cloning,andcharacterizationofproteins
involvedinintracellulartransportanditsregulation,aswellasindeciphering
correspondingmoleculareventswasachieved.Proteinsinvolvedinbudding,
fission,fusion,andsortinghavebeendiscovered,and,insomecases,apicture
ofhowsuchproteinsareassembledandworkhasbeenglimpsed.Incontrast,
perhaps surprisingly, a satisfactory understanding of how transport occurs
invivoattheorganellelevelhasnotbeenachieved,andthegeneralpicture
ofthisprocessremainsobscure.Asaconsequence,ourpresentviewofthe
overallmodeofintracellulartraffic(the“physiology”oftraffic)inlivingcells
isratherpoorlydeveloped.Althoughinvitroreconstitutionexperimentshave
beencrucialinestablishingtheminimumnumberofcomponentsrequired
forcarrierbuddingusingbiochemicaltechniques,amorecriticalevaluation
ofintracellulartransportcouldbecomplementedbyinvivoexperiments.
Thereasonsforthislagarebothtechnologicalandconceptual.Techno-
logical,becauseintracellulartrafficisanessentiallydynamiceventintimeand
2 * A.A.MironovandM.Pavelka
space.Yet,methodstostudyitsdynamicfeatureshavebeenlackinguntilvery
recently.Therecentadventofgreenfluorescentprotein(GFP)technologyhas
nowpartiallyfilledthisgap.Conceptual,becausethefieldhasbeendomi-
nateduntilveryrecentlybythemodelofintracellulartrafficbyanterograde
vesicles.Thevesicularschemehasbeenverysuccessfulinprovidingaframe-
worktointegrateanenormousamountofbiochemicalandgeneticevidence
collectedoverthepasttwodecadesintoacoherentpicture.
Inthepastyears,theprogressindevelopmentofnewscientificmethods
accelerated. New microscopic and biochemical methods, new scientific in-
strumentstogetherwiththedevelopmentofmoleculartoolshasgiventhe
possibilitytostudybiochemicalreactionsandmoleculesinsinglecellswitha
resolutionimpossibletoachievebefore.Significantprogresswasmadeinthe
development ofnewsynchronization protocols suitable forstudy oftrans-
portofseveralcargoes.
Ontheotherhand,thecompletedecipheringofthefullgeneticcodeof
yeast,humans,plantsandseveralotherspeciestogetherwiththedevelop-
ment of proteomics provided a significant number of new proteins and
details on protein–protein and protein–lipid interactions involved into the
traffic. In the literature, there are thousands of details regarding protein
interactions, their sorting signals, and the effects of their inhibition and
deletion.Assuch,itseemsthatthefieldofintracellulartransportappearsto
befacingaseriouscrisis.Therearenowsomanyproteinsandinter-protein
interactions involved that it has become almost impossible to follow and
understandthemeaningofallofthesedetails.
Themainideaofthisbookisthustocollectthefullrangeofexpertiseand
to examine the problems from differentpoints of view andwith different
approaches.
Thisbookisdevotedtothemolecularmechanismsofmorpho-functional
organizationoftheGAandsummarizesmostofnewdatarelatedtotheGA.
Thebookisacollectionofchapterswrittenbydifferentgroupsandtherefore
expressesdifferentviewsespeciallyonthemechanismsoftraffic.
Thepossibilitytofollowtheevaluationoftheintracellulartransportfrom
differentpointsofviewandonthebasisofdifferentexpertisescouldhelpto
resolvethiscontradictioninthefield.
There are several levels in the book. The main is the description of cell
physiology with emphasison the physiology ofintracellular transport. The
secondlevelisthepresentationofthemorphologyinwidetermincluding
light microscopy, analysis of live cells and so on. Finally, there are several
chaptersdevotedtothemolecularmechanismsinvolvedindifferentphysio-
logicalprocessesrelatedtointracellulartransport.Inthisbook,wecouldnot
excludecompletelysomedegreeofoverlapping.First,becausetheauthors
have different opinions about models of transport, and the second, it was
necessarytoillustratesomeideasfromdifferentpointsofview.Weapologize
to colleagues, whose relevant work has not been mentioned because of
spacelimitationsandfocusingonworkpublishedmostrecently.
Introduction * 3
Inthefirstpartofthebooktherearetwochapters.Oneisanoverviewof
thehistoryofthisfieldofcellbiology.Thesecondoneisamorpho-functional
overview of the main steps of intracellular transport including functional
organizationandarchitectureoftheGA.
In the second part of the book, main machineries operating along the
secretorypathwayarepresented.TheseincludeSNAREs,Rabs,COPII,COPI,
ARFs,ARFGEFsandARLs,COG,TRAPP,dynaminandcortactin,Golgienzymes
andsugartransporters,ERGIC-53,andotherluminallectins,GolginsandGolgi
matrix proteins. Lipids and lipid signalling, as well as common signalling
mechanismswillbeassessed.Additionally,theroleofcalciumandotherions
intheregulationofintracellulartransport,andtheroleofthecytoskeletonin
Golgifunctionwillbedescribed.
In the third part of the book, different transport steps of intracellular
transportwillbeevaluated,namely,ER-to-Golgitransport,intra-Golgitrans-
port,Golgi-to-PMtransport,Golgi-to-endosometransport,transportoflyso-
somal enzymes, retrograde endosome-to-TGN transport, and retrograde
PM-to-Golgitransport.Mechanismsofregulatedsecretionwillbeexplained
inaseparatechapterandinparticular,mechanismsinvolvedinformationof
mucingranulesaredescribed.Interactionsbetweenendocytosisandsecre-
torytransport,therelationshipbetweentheGAandcellpolarity,aswellas
structure and domain organization of the trans-Golgi network, and the
questions of GA formation and inheritance will be discussed in separate
chapters.
Finally, in the last part of the book the reader will find some aspects
about the peculiarities of intracellular transport in different organisms:
plants,yeast,andprotists.Aseparatechapterisdevotedtotheendomem-
brane ultrastructure and dynamics in yeasts. Finally, the models of Golgi
evolutionwillbediscussedinthefinalchapterofthispart.
Thisbookisintendedforcellbiologistsandhistologists,whoworkwith
students,andalsoforscientistsworkinginotherfieldsofbiologyaswellas
forstudentsperse.Themostimportantitemforteachingistheunderstand-
ing of not a single or several mechanisms but the comprehensive view
uponthefulldramaofdevelopmentofscientificmodels.Thereadersshould
understand the logic of model replacements. Therefore, the aim of the
bookistomakethefieldofintracellulartransportmoreunderstandableby
keepingitassimpleaspossible,butalsoasfullaspossible,andatminimalcost.
Finally,wewouldliketomentionthequestionofterminology.Sincethe
originaltermwas“Golgiapparatus”,thistermisusedinmostofthechapters.
However, both terms, Golgi apparatus and Golgi complex, are customary
today.
General considerations