Characterization of Beclin 1 and USP14 Interactions

Myc-tagged USP14, Myc-tagged Beclin 1, Flag-tagged USP14, Flag-tagged TRAF6, HA-tagged TAB 2, HA-tagged Ub, and Myc-tagged Ub vectors were used in this study. Myc-tagged Beclin 1 truncated mutants, Myc-tagged Beclin 1 1-269, and Myc-tagged Beclin 1 1-127 were generated using specific primers shown in Supplementary Material (Table S1 in Supplementary Material). HA-tagged TAB 2 truncated mutants, HA-tagged TAB 2 ΔCUE, HA-tagged TAB 2 1-518, and HA-tagged TAB 2 518-693 were generated with specific primers shown in Supplementary Material (Table S2 in Supplementary Material). Flag-tagged TRAF6 truncated mutants, Flag-tagged TRAF6 110-522, Flag-tagged TRAF6 260-522, and Flag-tagged TRAF6 349-522 were generated as described previously (33 (link)). Flag-tagged Beclin 1 vector was purchased from Addgene (Cambridge, MA, USA). Myc-tagged Beclin 1 vector was generated by PCR using Flag-tagged Beclin 1 as a template and inserted into pCMV-Myc (Addgene). pCDNA3-USP14 vector was kindly provided by Dr. M. J. Lee (Seoul National University College of Medicine, Seoul, Korea). Myc-tagged USP14 or Flag-tagged USP14 vector was generated by PCR using pCDNA3-USP14 as a template.

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Min Y., Lee S., Kim M.J., Chun E, & Lee K.Y. (2017). Ubiquitin-Specific Protease 14 Negatively Regulates Toll-Like Receptor 4-Mediated Signaling and Autophagy Induction by Inhibiting Ubiquitination of TAK1-Binding Protein 2 and Beclin 1. Frontiers in Immunology, 8, 1827.

Publication 2017

pCMV-Myc

Beclin 1 Medicine Primers Traf6 Usp14 Vector

Corresponding Organization :

Other organizations : Sungkyunkwan University, Samsung (South Korea), Harvard University, Samsung Medical Center

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Protocol cited in 4 other protocols

Variable analysis

independent variables

Myc-tagged USP14
Myc-tagged Beclin 1
Flag-tagged USP14
Flag-tagged TRAF6
HA-tagged TAB 2
HA-tagged Ub
Myc-tagged Ub
Myc-tagged Beclin 1 truncated mutants (Myc-tagged Beclin 1 1-269, Myc-tagged Beclin 1 1-127)
HA-tagged TAB 2 truncated mutants (HA-tagged TAB 2 ΔCUE, HA-tagged TAB 2 1-518, HA-tagged TAB 2 518-693)
Flag-tagged TRAF6 truncated mutants (Flag-tagged TRAF6 110-522, Flag-tagged TRAF6 260-522, Flag-tagged TRAF6 349-522)

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

positive controls

Flag-tagged Beclin 1 vector purchased from Addgene
PCDNA3-USP14 vector provided by Dr. M. J. Lee

negative controls

Not mentioned

Annotations

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