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Identification of Nuclear Localization Sequence of G Protein-Coupled Receptor Kinase 6 under Depalmitoylation
Huang Xishi1, Li Ruirui1, Huang Lanzhen1,2, Yang Jing1,2, Wu Qunying1,3, Jiang Xiaoshan1,4*
1Cell Signaling Laborary, Guilin Medical University, Guilin 541004, China; 2Center for Science Research, Guilin Medical University, Guilin 541004, China; 3Department of Genetics and Cell Biology, Guilin Medical University, Guilin 541004, China; 4Graduate College, Guilin Medical University, Guilin 541004, China
Abstract: G protein-coupled receptor kinase 6 (GRK6) is capable of achieving nuclear translocation through depalmitoylation. Although a nuclear localization sequence (NLS) region has reported， the key residue(s) in the NLS domain as well as its effect on the kinase activity are not yet clear. In this study， we first verified a predicted NLS domain through screening a series of GRK6 deletion mutants under the condition of depalmitoylation. Then， we further identified three residues， Lys(K)389， Lys(K)390 and Lys(K)391， which respectively played essential role in the nuclear translocalization of GRK6 by using a point mutation technology. Finally， the biological function of the mutants in the M3 receptor signaling， comparing to GRK6 WT， was analyzed using a calcium mobilization assay and found no significant effect on their kinase activities. This study provides valuable information to further study the nuclear translocation mechanism and function of GRK6.
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