Novel export and import pathways in S. cerevisiae identified by an engineered SUMO system

Vera Rodriguez, Arturo

dc.contributor.advisor	Görlich, Dirk Prof. Dr.
dc.contributor.author	Vera Rodriguez, Arturo
dc.date.accessioned	2018-09-07T08:28:16Z
dc.date.available	2018-09-07T08:28:16Z
dc.date.issued	2018-09-07
dc.identifier.uri	http://hdl.handle.net/11858/00-1735-0000-002E-E49F-0
dc.identifier.uri	http://dx.doi.org/10.53846/goediss-7045
dc.language.iso	eng	de
dc.rights.uri	http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject.ddc	570	de
dc.title	Novel export and import pathways in S. cerevisiae identified by an engineered SUMO system	de
dc.type	doctoralThesis	de
dc.contributor.referee	Rodnina, Marina Prof. Dr.
dc.date.examination	2017-06-26
dc.description.abstracteng	Nuclear transport receptors (NTRs) mediate the translocation of cargos between the cytoplasm and the nucleus. NTRs are classified either as importins or exportins according to the directionality of the cargo transport. To identify new interaction partners of so far poorly characterized NTRs from S. cerevisiae, we developed a novel protein purification approach based on an engineered SUMO-specific protease (SUMOvera protease) and its specific SUMO substrate (SUMOvera). SUMOvera can be used as a stable fusion tag in virtually all eukaryotic systems since it is not recognized by SUMO-specific proteases from yeast, plant, human, amphibian, insect and protozoa. In addition, the SUMOvera protease can be over-expressed in S. cerevisiae without causing toxicity unlike yeast and B. distachyon SUMO-specific proteases. The SUMOvera system has also an orthogonal specificity to the SUMOstar/SUMOstar protease pair to allow the purification of protein complexes with a defined stoichiometry in eukaryotic hosts. Using the SUMOvera system, we discovered that yeast Lph2 and Pdr6 mediate the nuclear export and import of different cargos. Specifically, we showed that Lph2 exports and binds to the translation initiation factor eIF4A in a Ran-GTP-dependent manner, and that Pdr6 recognizes Ubc9 as an import substrate as well as eEF2 and eIF5A as export cargos. Lph2 and Pdr6 have been only described as importins; however, our data indicate that they act as bidirectional NTRs that shuttle distinct sets of cargos in opposite directions through the nuclear envelope. Overall, we describe two novel bidirectional NTRs in S. cerevisiae in addition to Msn5. The reported findings suggest that bidirectionality in NTRs might be more common than previously assumed, and that there might be other bidirectional NTRs in different organisms that still need to be identified.	de
dc.contributor.coReferee	Urlaub, Henning Prof. Dr.
dc.contributor.thirdReferee	Klopfenstein, Dieter Dr.
dc.contributor.thirdReferee	Shcherbata, Halyna PD Dr.
dc.contributor.thirdReferee	Dosch, Roland Dr.
dc.subject.eng	SUMO	de
dc.subject.eng	NTRs	de
dc.subject.eng	S. cerevisiae	de
dc.subject.eng	Pdr6	de
dc.subject.eng	Lph2	de
dc.subject.eng	Ubc9	de
dc.subject.eng	eIF5A	de
dc.subject.eng	eEF2	de
dc.subject.eng	eIF4A	de
dc.identifier.urn	urn:nbn:de:gbv:7-11858/00-1735-0000-002E-E49F-0-6
dc.affiliation.institute	Göttinger Graduiertenschule für Neurowissenschaften, Biophysik und molekulare Biowissenschaften (GGNB)	de
dc.subject.gokfull	Biologie (PPN619462639)	de
dc.identifier.ppn	1030476101

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Das Dokument erscheint in:

GGNB - Göttinger Graduiertenzentrum für Neurowissenschaften, Biophysik und molekulare Biowissenschaften [1115]
GGNB - Göttingen Graduate Center for Neurosciences, Biophysics and Molecular Biosciences

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