Universität zu Köln

Song, Lin, Rijal, Ramesh ORCID: 0000-0003-0498-1064, Karow, Matte, Stumpf, Maria, Hahn, Oliver ORCID: 0000-0001-9015-4545, Park, Laura, Insall, Robert ORCID: 0000-0003-4898-040X, Schroeder, Rolf, Hofmann, Andreas ORCID: 0000-0003-4408-5467, Clemen, Christoph S. and Eichinger, Ludwig ORCID: 0000-0003-1594-6117 (2018). Expression of N471D strumpellin leads to defects in the endolysosomal system. Dis. Model. Mech., 11 (9). CAMBRIDGE: COMPANY BIOLOGISTS LTD. ISSN 1754-8411

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Abstract

Hereditary spastic paraplegias (HSPs) are genetically diverse and clinically characterised by lower limb weakness and spasticity. The N471 D and several other point mutations of human strumpellin (Str; also known as WASHC5), a member of the Wiskott-Aldrich syndrome protein and SCAR homologue (WASH) complex, have been shown to cause a form of HSP known as spastic paraplegia 8 (SPG8). To investigate the molecular functions of wild-type (WT) and N417D Str, we generated Dictyostelium Str(-) cells and ectopically expressed Str(WT-)GFP or Str(N471D)-GFP in Str(- )and WT cells. Overexpression of both proteins apparently caused a defect in cell division, as we observed a clear increase in multinucleate cells. Real-time PCR analyses revealed no transcriptional changes in WASH complex subunits in Str(-) cells, but western blots showed a twofold decrease in the SWIP subunit. GFP-trap experiments in conjunction with mass-spectrometric analysis revealed many previously known, as well as new, Str-interacting proteins, and also proteins that no longer bind to Str(N4471D). At the cellular level, Str(-) cells displayed defects in cell growth, phagocytosis, macropinocytosis, exocytosis and lysosomal function. Expression of Str(WT-)GFP in St-cells rescued all observed defects. In contrast, expression of Str(N471D)-GFP could not rescue lysosome morphology and exocytosis of indigestible material. Our results underscore a key role for the WASH complex and its core subunit, Str, in the endolysosomal system, and highlight the fundamental importance of the Str N471 residue for maintaining lysosome morphology and dynamics. Our data indicate that the SPG8-causing N471D mutation leads to a partial loss of Str function in the endolysosomal system. This article has an associated First Person interview with the first author of the paper.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Song, Lin UNSPECIFIED UNSPECIFIED UNSPECIFIED Rijal, Ramesh UNSPECIFIED orcid.org/0000-0003-0498-1064 UNSPECIFIED Karow, Matte UNSPECIFIED UNSPECIFIED UNSPECIFIED Stumpf, Maria UNSPECIFIED UNSPECIFIED UNSPECIFIED Hahn, Oliver UNSPECIFIED orcid.org/0000-0001-9015-4545 UNSPECIFIED Park, Laura UNSPECIFIED UNSPECIFIED UNSPECIFIED Insall, Robert UNSPECIFIED orcid.org/0000-0003-4898-040X UNSPECIFIED Schroeder, Rolf UNSPECIFIED UNSPECIFIED UNSPECIFIED Hofmann, Andreas UNSPECIFIED orcid.org/0000-0003-4408-5467 UNSPECIFIED Clemen, Christoph S. UNSPECIFIED UNSPECIFIED UNSPECIFIED Eichinger, Ludwig UNSPECIFIED orcid.org/0000-0003-1594-6117 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-174292
DOI:	10.1242/dmm.033449
Journal or Publication Title:	Dis. Model. Mech.
Volume:	11
Number:	9
Date:	2018
Publisher:	COMPANY BIOLOGISTS LTD
Place of Publication:	CAMBRIDGE
ISSN:	1754-8411
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language HEREDITARY SPASTIC PARAPLEGIA; WASH COMPLEX; DICTYOSTELIUM-DISCOIDEUM; ACTIN POLYMERIZATION; PROTEIN AGGREGATION; AUTOPHAGY; MEMBRANE; MUTATION; PATHWAY; DISEASE Multiple languages Cell Biology; Pathology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/17429