FIGURE

Fig. 3

ID
ZDB-FIG-231004-7
Publication
Yong et al., 2023 - Cryo-EM structure of the Mon1-Ccz1-RMC1 complex reveals molecular basis of metazoan RAB7A activation
Other Figures
All Figure Page
Back to All Figure Page
Fig. 3

The assembly of RMC1 with Mon1 and Ccz1. (A) Detailed interactions of the interfaces between DmRMC1 (pink) and DmCcz1 (green). Secondary structures of DmRMC1 and DmCcz1 are labeled as indicated. (BE) Shown are hotspots1–4, with residues critical for stabilizing the interface shown in sticks. The rotation angles are shown as indicated. (F) HEK293T cells were transfected with GFP-tagged RMC1 WT, RMC1HQY/AAA or RMC1QDR/ARE mutants and subjected to GFP-nanotrap for immunoprecipitation. RMC1 and endogenous Ccz1, Mon1B were detected via antibodies against GFP, Ccz1 and Mon1B, respectively. (G) HEK293T cells were cotransfected with GFP-tagged RMC1 and Flag-tagged Ccz1 WT or Ccz1NL/AA mutant, and then subjected to Flag-nanotrap for immunoprecipitation. RMC1 and Ccz1 were detected via antibodies against GFP and Flag, respectively. (H) Detailed interactions of the interfaces between DmRMC1 (pink) and DmMon1 (blue). Secondary structures of DmRMC1 and DmMon1 are labeled as indicated. (IK) Shown are interfaces between DmRMC1 (pink) and DmMon1 (blue), with residues critical for stabilizing the interface shown in sticks. The rotation angle are shown as indicated. (L) HEK293T cells were transfected with GFP-tagged RMC1 WT, RMC1LRR/AEE or RMC1DY/RA mutants and subjected to GFP-nanotrap for immunoprecipitation. RMC1 and endogenous Mon1B were detected via antibodies against GFP and Mon1B, respectively. (M) HEK293T cells were cotransfected with GFP-tagged RMC1 and Flag-tagged Mon1B WT or Mon1BSL/AA mutant, and then subjected to Flag-nanotrap for immunoprecipitation. RMC1 and Mon1B were detected via antibodies against GFP and Flag, respectively. Black dashed lines indicate hydrogen bonds or salt bridges.

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Proc. Natl. Acad. Sci. USA