Cytoplasmic Transport Machinery of the SPF27 Homologue Num1 in Ustilago maydis

Scientific Reports
Lu ZhouJörg Kämper

Abstract

In the phytopathogenic basidiomycete Ustilago maydis, the Num1 protein has a pivotal function in hyphal morphogenesis. Num1 functions as a core component of the spliceosome-associated Prp19/CDC5 complex (NTC). The interaction of Num1 with the kinesin motor Kin1 suggests a connection between a component of the splicing machinery and cytoplasmic trafficking processes. Previously it was shown that Num1 localizes predominantly in the nucleus; however, due to the diffraction-limited spatial resolution of conventional optical microscopy, it was not possible to attribute the localization to specific structures within the cytoplasm. We have now employed super-resolution localization microscopy to visualize Num1 in the cytoplasm by fusing it to a tandem dimeric Eos fluorescent protein (tdEosFP). The Num1 protein is localized within the cytoplasm with an enhanced density in the vicinity of microtubules. Num1 movement is found predominantly close to the nucleus. Movement is dependent on its interaction partner Kin1, but independent of Kin3. Our results provide strong evidence that, in addition to its involvement in splicing in the nucleus, Num1 has an additional functional role in the cytosol connected to the Kin1 motor protein.

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Citations

Jan 29, 2019·Current Microbiology·Dario R Olicón-HernándezGuadalupe Guerra-Sánchez
Jul 13, 2019·Biochemical Society Transactions·Ilijana VojnovicUlrike Endesfelder
Apr 9, 2020·Molecular Biology of the Cell·Rongde QiuXin Xiang
Aug 18, 2021·EMBO Reports·Kira MüntjesMichael Feldbrügge
Nov 25, 2021·Molecular Microbiology·Karina SchneiderJörg Kämper

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Datasets Mentioned

BETA
AAV54099.1

Methods Mentioned

BETA
two-hybrid
imaging technique
super-resolution
fluorescence microscopy
Confocal microscopy
Localization
super-resolution microscopy
PCR
Fluorescence

Software Mentioned

livePALM
ImageJ
MATLAB

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