title: Identification of MANF as a protein interacting with RTN1-C.
abstract: Reticulons (RTNs) constitute a family of endoplasmic reticulum (ER)-associated proteins with a reticular distribution. Recently, evidence has shown that they exert a cancer-speciﬁc proapoptotic function via interaction or modulation of speciﬁc proteins. Such evidence is particularly associated with the RTN1-C family members. In order to explore proteins that interact with RTN1-C, the yeast two-hybrid system and regular molecular biological techniques were used to screen the human fetal brain cDNA library. As a result, seven RTN1-C interacting proteins including Homo sapiens mesencephalic astrocyte-derived neurotrophic factor (MANF) were obtained. The interactions between RTN1-C and its interacting proteins were confirmed by β-galactosidase assay and growth test in selective media. Moreover, the MANF/RTN1-C interaction was verified in vitro by glutathione S-transferase pull-down assay and in vivo by immunoprecipitation assay. By immunoﬂuorescence assay, it was found that MANF co-localized with RTN1-C in the ER. Knockdown of RTN1-C reduced the localization of MANF in the ER. These results provide clues to further explore the function of RTN1-C and MANF in neurodegenerative diseases and cancer.
authors.affiliation: School of Pharmacy, Anhui Medical University, Hefei 230032, China Department of Anaesthesiology of the First Afﬁliated Hospital, Anhui Medical University, Hefei 230032, China.
authors.forename: Lijian
authors.initials: L
authors.lastname: Chen
authors.affiliation: School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China.
authors.forename: Lijuan
authors.initials: L
authors.lastname: Wan
authors.affiliation: School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China dujane@163.com shenyx@ahmu.edu.cn.
authors.forename: Jian
authors.initials: J
authors.lastname: Du
authors.affiliation: School of Pharmacy, Anhui Medical University, Hefei 230032, China School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China dujane@163.com shenyx@ahmu.edu.cn.
authors.forename: Yuxian
authors.initials: Y
authors.lastname: Shen
mesh_terms: D001921:Brain / Q000378:metabolism; D045744:Cell Line, Tumor; D018076:DNA, Complementary / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D008856:Microscopy, Fluorescence; D009414:Nerve Growth Factors / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D019636:Neurodegenerative Diseases / Q000378:metabolism; D011485:Protein Binding; D025941:Protein Interaction Mapping; D011994:Recombinant Proteins / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; C099695:MANF protein, human; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; C081781:RTN1 protein, human; D011994:Recombinant Proteins; D005982:Glutathione Transferase; D001616:beta-Galactosidase
keywords: MANF; RTN1-C; protein interaction; reticulon; yeast two-hybrid