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Activities of Pex19p to interact with <jats:styled-content style=\"fixed-case\">PMPs<\/jats:styled-content> on one hand and Pex3p on the other, including formation of ternary complexes between Pex19p, <jats:styled-content style=\"fixed-case\">PMP<\/jats:styled-content> and Pex3p, strongly support posttranslational translocation of <jats:styled-content style=\"fixed-case\">PMPs<\/jats:styled-content> via the Pex19p\u2010 and Pex3p\u2010dependent direct pathway, termed the class I pathway. However, it remains elusive whether Pex19p\u2010<jats:styled-content style=\"fixed-case\">PMP<\/jats:styled-content> complexes are indeed capable of being imported into peroxisomal membranes through the interaction between Pex19p and Pex3p. We resolve this issue by investigating the targeting process of several topologically distinct <jats:styled-content style=\"fixed-case\">PMPs<\/jats:styled-content>, including multimembrane spanning <jats:styled-content style=\"fixed-case\">PMPs<\/jats:styled-content>. We show here that Pex19p forms cytosolic complexes with <jats:styled-content style=\"fixed-case\">PMPs<\/jats:styled-content> and directly translocates them to peroxisomes. Using a semi\u2010intact mammalian cell\u2010based import assay system, we prove that <jats:styled-content style=\"fixed-case\">PMPs<\/jats:styled-content> in the cytosolic complexes are imported into peroxisomes via the interaction between cargo\u2010loaded Pex19p and Pex3p. Furthermore, we demonstrate for the first time that peroxisomal targeting of <jats:styled-content style=\"fixed-case\">ATAD1<\/jats:styled-content>, an N\u2010terminally signal\u2010anchored protein that resides on both mitochondria and peroxisomes, is also achieved through the Pex19p\u2010 and Pex3p\u2010dependent class I pathway. Together, our results suggest that translocation of <jats:styled-content style=\"fixed-case\">PMPs<\/jats:styled-content> via the class I pathway is a common event in mammalian cells.<\/jats:p><jats:p><jats:inline-graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"graphic\/tra12376-gra-0001.png\" xlink:title=\"image\"\/><\/jats:p>","DOI":"10.1111\/tra.12376","type":"journal-article","created":{"date-parts":[[2016,1,18]],"date-time":"2016-01-18T13:18:14Z","timestamp":1453123094000},"page":"433-455","source":"Crossref","is-referenced-by-count":33,"title":["Assembly of Peroxisomal Membrane Proteins via the Direct Pex19p\u2010Pex3p Pathway"],"prefix":"10.1111","volume":"17","author":[{"given":"Yuqiong","family":"Liu","sequence":"first","affiliation":[{"name":"Graduate School of Systems Life Sciences Kyushu University Graduate School  Nishi\u2010ku Fukuoka 819\u20100395 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