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A nanobody suite for yeast scaffold nucleoporins provides details of the nuclear pore complex structure.


ABSTRACT: Nuclear pore complexes (NPCs) are the main conduits for molecular exchange across the nuclear envelope. The NPC is a modular assembly of ~500 individual proteins, called nucleoporins or nups. Most scaffolding nups are organized in two multimeric subcomplexes, the Nup84 or Y complex and the Nic96 or inner ring complex. Working in S. cerevisiae, and to study the assembly of these two essential subcomplexes, we here develop a set of twelve nanobodies that recognize seven constituent nucleoporins of the Y and Nic96 complexes. These nanobodies all bind specifically and with high affinity. We present structures of several nup-nanobody complexes, revealing their binding sites. Additionally, constitutive expression of the nanobody suite in S. cerevisiae detect accessible and obstructed surfaces of the Y complex and Nic96 within the NPC. Overall, this suite of nanobodies provides a unique and versatile toolkit for the study of the NPC.

SUBMITTER: Nordeen SA 

PROVIDER: S-EPMC7710722 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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A nanobody suite for yeast scaffold nucleoporins provides details of the nuclear pore complex structure.

Nordeen Sarah A SA   Andersen Kasper R KR   Knockenhauer Kevin E KE   Ingram Jessica R JR   Ploegh Hidde L HL   Schwartz Thomas U TU  

Nature communications 20201202 1


Nuclear pore complexes (NPCs) are the main conduits for molecular exchange across the nuclear envelope. The NPC is a modular assembly of ~500 individual proteins, called nucleoporins or nups. Most scaffolding nups are organized in two multimeric subcomplexes, the Nup84 or Y complex and the Nic96 or inner ring complex. Working in S. cerevisiae, and to study the assembly of these two essential subcomplexes, we here develop a set of twelve nanobodies that recognize seven constituent nucleoporins of  ...[more]

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