期刊
ACTA BIOCHIMICA ET BIOPHYSICA SINICA
卷 47, 期 2, 页码 91-97出版社
OXFORD UNIV PRESS
DOI: 10.1093/abbs/gmu125
关键词
reticulon; RTN1-C; MANF; yeast two-hybrid; protein interaction
资金
- National Natural Science Foundation of China [81271864, 30801329]
- Fok Ying Tung College Young Teachers Fund of the Chinese Ministry of Education [131033]
- Distinguished Young Scholar of Anhui Province [10040606Y19, 1308085JGD11]
- Natural Science Foundation of Anhui Province, China [1308085MH118]
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 beta-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.
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