title: A new functional screening system for identification of regulators for the generation of amyloid beta-protein.
abstract: Presenilin (PS) is essential for gamma-cleavage, which is required for the generation of amyloid beta-protein (Abeta) from the beta-amyloid precursor protein. However, it remains to be clarified how gamma-cleavage is regulated. To elucidate the regulation of PS-mediated gamma-cleavage, we developed a new functional screening method for identifying cDNA that enhances gamma-cleavage. This screening system utilizes our own developed cell line, where the expression of cDNA that enhances gamma-cleavage confers puromycin resistance. The cDNA library is retrovirally delivered to the above-mentioned cell line, allowing the identification of our target cDNAs by a combination of puromycin resistance selection and Abeta assay screening. With this screening method, we isolated several cDNAs enhancing gamma-cleavage, including the previously reported Herp. Here we also demonstrate that Rab1A, identified with this screening, can be a regulator of Abeta generation. Thus, our established screening method is a powerful tool for identifying multiple regulators involved in gamma-cleavage in the Abeta generation pathway, including modulators of gamma-secretase activity or the intracellular trafficking of factors necessary for gamma-cleavage.
authors.affiliation: Department of Dementia Research, National Institute for Longevity Sciences, 36-3 Gengo, Morioka, Obu, Aichi 474-8522, Japan. hkomano@nils.go.jp
authors.forename: Hiroto
authors.initials: H
authors.lastname: Komano
authors.forename: Hirohisa
authors.initials: H
authors.lastname: Shiraishi
authors.forename: Yuuki
authors.initials: Y
authors.lastname: Kawamura
authors.forename: Xiaorei
authors.initials: X
authors.lastname: Sai
authors.forename: Ryo
authors.initials: R
authors.lastname: Suzuki
authors.forename: Lutgarde
authors.initials: L
authors.lastname: Serneels
authors.forename: Masashi
authors.initials: M
authors.lastname: Kawaichi
authors.forename: Toshio
authors.initials: T
authors.lastname: Kitamura
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Yanagisawa
mesh_terms: D053829:Amyloid Precursor Protein Secretases; D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000378:metabolism*; D000818:Animals; D016282:Aspartic Acid Endopeptidases; D051792:Basic Helix-Loop-Helix Proteins; D002460:Cell Line; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004351:Drug Resistance; D010450:Endopeptidases / Q000378:metabolism; D015723:Gene Library; D005821:Genetic Techniques; D006624:Hippocampus / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D015151:Immunoblotting; D051379:Mice; D018345:Mice, Knockout; D008957:Models, Genetic; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D011485:Protein Binding; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011691:Puromycin / Q000494:pharmacology; D012190:Retroviridae / Q000235:genetics; D000072056:Transcription Factor HES-1; D020761:rab1 GTP-Binding Proteins / Q000378:metabolism
chemical_list: D053829:Amyloid Precursor Protein Secretases; D016229:Amyloid beta-Peptides; D016282:Aspartic Acid Endopeptidases; D051792:Basic Helix-Loop-Helix Proteins; D018076:DNA, Complementary; D010450:Endopeptidases; D018398:Homeodomain Proteins; D011500:Protein Synthesis Inhibitors; D011691:Puromycin; D000072056:Transcription Factor HES-1; D020761:rab1 GTP-Binding Proteins; C489734:Hes1 protein, mouse; C078493:HES1 protein, human; C509280:BACE1 protein, human; C509282:Bace1 protein, mouse