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Ubiquitin-conjugating activity by PEX4 is required for efficient protein transport to peroxisomes in Arabidopsis thaliana 1 Results 2 Decrease of efficiency of PTS1- and PTS2-dependent protein transport in the apem7 mutant 2 Peroxisomal functions in apem7 mutants 2 The APEM7 gene encodes PEX4, a member of the UBC family 3 PEX4 is localized on peroxisomal membranes 3 PEX4 is ubiquitinated, and the apem7 mutation disturbs the ubiquitination of PEX4 in vivo 3 Abnormal localization of PEX5 in apem7 mutants 6 PEX4 is expressed in various tissues 6 Discussion 6 Effects of the apem7/pex4 mutation on protein transport to peroxisomes 7 Detection of UBC activity and the effect of PEX4 ubiquitination on protein transport 7 Experimental procedures 8 Plant materials 8 Confocal microscopic observation 8 Map-based cloning of the APEM7 locus 8 Generation of transgenic plants 8 Preparation of antibodies against PEX4 9 Immunoblot analysis 9 Electron microscopic analysis 9 In vitro transcription/translation for ubiquitination 9 Data availability 10 Supporting information 10 Author contributions 10 Funding and additional information 10 References 10
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