Posted On: 01/05/2015 10:30:07 AM
Post# of 30066
MANF>CANCER
Identification of MANF as a protein interacting with RTN1-CReticulons (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 pertinent for the RTN1-C family member. In order to identify 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 using both β-galactosidase assay and growth test in selective media. Moreover, the MANF/RTN1-C interaction was confirmed in vitro by glutathione S-transferase pull-down and in vivo by immunoprecipitation assays. 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.
Identification of MANF as a protein interacting with RTN1-CReticulons (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 pertinent for the RTN1-C family member. In order to identify 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 using both β-galactosidase assay and growth test in selective media. Moreover, the MANF/RTN1-C interaction was confirmed in vitro by glutathione S-transferase pull-down and in vivo by immunoprecipitation assays. 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.
(0)
(0)
