Biochemical and Biophysical Research Communications
Function and subcellular location of Ro52β
Section snippets
Materials and methods
Cell culture. Human embryonic kidney HEK293 cells and HEK293T cells (American Type Culture Collection, Manassas, VA) were maintained in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal calf serum and antibiotics.
Antibodies. Mouse anti-FLAG antibody (M5) was purchased from Sigma Chemical Company (St. Louis, MO). Mouse anti-RH antibody (specific for the amino acid sequences RGSHHHH and GGSHHHH) was purchased from QIAGEN (Santa Clara, CA). Mouse anti-HA antibody (16B12) was purchased
Ro52β structurally lacks the leucine zipper
To compare human Ro52α and Ro52β, we first summarized their domain structure, which was previously reported [2]. As shown in Fig. 1, Ro52α possesses a RING finger and a B box at the N-terminal region. This RING-finger motif is required for the activity of E3 ubiquitin ligase [3]. In the central domain, Ro52α possesses two coiled coils, the second of which is a leucine zipper encoded in exon 4. Recently, Ro52α was shown to form homodimers through its leucine zipper [11]. At the C-terminus, Ro52α
Acknowledgment
This work was supported by National Institutes of Health Grant R01 DK56298 (to T.K.).
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