Human Stefin B Role in Cell's Response to Misfolded Proteins and Autophagy
Polajnar, Mira (Jožef Stefan International Postgraduate School)
Zavašnik-Bergant, Tina (Jožef Stefan Institute. Department of Biochemistry and Molecular and Structural Biology)
Škerget, Katja (Jožef Stefan Institute. Department of Molecular and Biomedical Science)
Vizovišek, Matej (Jožef Stefan International Postgraduate School)
Vidmar, Robert (Jožef Stefan International Postgraduate School)
Fonović, Marko (Jožef Stefan International Postgraduate School)
Kopitar-Jerala, Nataša (Jožef Stefan Institute. Department of Biochemistry and Molecular and Structural Biology)
Petrovič, Uroš (Jožef Stefan Institute. Department of Molecular and Biomedical Science)
Navarro, Susanna (Universitat Autònoma de Barcelona. Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")
Ventura, Salvador (Universitat Autònoma de Barcelona. Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")
Žerovnik, Eva (Center of Excellence for integrated approaches in chemistry and biology of proteins)

Data:	2014
Resum:	Alternative functions, apart from cathepsins inhibition, are being discovered for stefin B. Here, we investigate its role in vesicular trafficking and autophagy. Astrocytes isolated from stefin B knock-out (KO) mice exhibited an increased level of protein aggregates scattered throughout the cytoplasm. Addition of stefin B monomers or small oligomers to the cell medium reverted this phenotype, as imaged by confocal microscopy. To monitor the identity of proteins embedded within aggregates in wild type (wt) and KO cells, the insoluble cell lysate fractions were isolated and analyzed by mass spectrometry. Chaperones, tubulins, dyneins, and proteosomal components were detected in the insoluble fraction of wt cells but not in KO aggregates. In contrast, the insoluble fraction of KO cells exhibited increased levels of apolipoprotein E, fibronectin, clusterin, major prion protein, and serpins H1 and I2 and some proteins of lysosomal origin, such as cathepsin D and CD63, relative to wt astrocytes. Analysis of autophagy activity demonstrated that this pathway was less functional in KO astrocytes. In addition, synthetic dosage lethality (SDL) gene interactions analysis in Saccharomyces cerevisiae expressing human stefin B suggests a role in transport of vesicles and vacuoles These activities would contribute, directly or indirectly to completion of autophagy in wt astrocytes and would account for the accumulation of protein aggregates in KO cells, since autophagy is a key pathway for the clearance of intracellular protein aggregates.
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Publicat a:	PloS one, Vol. 9, Issue 7 (July 2014) , art. e102500, ISSN 1932-6203

DOI: 10.1371/journal.pone.0102500
PMID: 25047918

15 p, 4.6 MB

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