Clathrin Heavy Chain Knockdown Impacts CXCR4 Signaling and Post-translational Modification
Recent research has implicated endocytic pathways as important regulators of receptor signaling. However, the role of endocytosis in regulating chemokine CXC receptor 4 (CXCR4) signaling remains largely unknown. In the present work we systematically investigate the impact of clathrin knockdown on CX...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2019-05-01
|
| Series: | Frontiers in Cell and Developmental Biology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fcell.2019.00077/full |
| _version_ | 1811221396198522880 |
|---|---|
| author | Maxwell S. DeNies Luciana K. Rosselli-Murai Santiago Schnell Santiago Schnell Santiago Schnell Allen P. Liu Allen P. Liu Allen P. Liu Allen P. Liu |
| author_facet | Maxwell S. DeNies Luciana K. Rosselli-Murai Santiago Schnell Santiago Schnell Santiago Schnell Allen P. Liu Allen P. Liu Allen P. Liu Allen P. Liu |
| author_sort | Maxwell S. DeNies |
| collection | DOAJ |
| description | Recent research has implicated endocytic pathways as important regulators of receptor signaling. However, the role of endocytosis in regulating chemokine CXC receptor 4 (CXCR4) signaling remains largely unknown. In the present work we systematically investigate the impact of clathrin knockdown on CXCR4 internalization, signaling, and receptor post-translational modification. Inhibition of clathrin-mediated endocytosis (CME) significantly reduced CXCR4 internalization. In contrast to other receptors, clathrin knockdown increased CXCL12-dependent ERK1/2 signaling. Simultaneous inhibition of CME and lipid raft disruption abrogated this increase in ERK1/2 phosphorylation suggesting that endocytic pathway compensation can influence signaling outcomes. Interestingly, using an antibody sensitive to CXCR4 post-translational modification, we also found that our ability to detect CXCR4 was drastically reduced upon clathrin knockdown. We hypothesize that this effect was due to differences in receptor post-translational modification as total CXCR4 protein and mRNA levels were unchanged. Lastly, we show that clathrin knockdown reduced CXCL12-dependent cell migration irrespective of an observed increase in ERK1/2 phosphorylation. Altogether, this work supports a complex model by which modulation of endocytosis affects not only receptor signaling and internalization but also receptor post-translational modification. |
| first_indexed | 2024-04-12T07:58:31Z |
| format | Article |
| id | doaj.art-3ab3e75fab9e4779825f6991b4d33288 |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-04-12T07:58:31Z |
| publishDate | 2019-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-3ab3e75fab9e4779825f6991b4d332882022-12-22T03:41:24ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2019-05-01710.3389/fcell.2019.00077443268Clathrin Heavy Chain Knockdown Impacts CXCR4 Signaling and Post-translational ModificationMaxwell S. DeNies0Luciana K. Rosselli-Murai1Santiago Schnell2Santiago Schnell3Santiago Schnell4Allen P. Liu5Allen P. Liu6Allen P. Liu7Allen P. Liu8Cellular and Molecular Biology Graduate Program, University of Michigan Medical School, Ann Arbor, MI, United StatesDepartment of Pharmacology, University of Michigan Medical School, Ann Arbor, MI, United StatesCellular and Molecular Biology Graduate Program, University of Michigan Medical School, Ann Arbor, MI, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan Medical School, Ann Arbor, MI, United StatesDepartment of Computational Medicine and Bioinformatics, University of Michigan Medical School, Ann Arbor, MI, United StatesCellular and Molecular Biology Graduate Program, University of Michigan Medical School, Ann Arbor, MI, United StatesDepartment of Mechanical Engineering, University of Michigan, Ann Arbor, MI, United StatesDepartment of Biomedical Engineering, University of Michigan, Ann Arbor, MI, United StatesDepartment of Biophysics, University of Michigan, Ann Arbor, MI, United StatesRecent research has implicated endocytic pathways as important regulators of receptor signaling. However, the role of endocytosis in regulating chemokine CXC receptor 4 (CXCR4) signaling remains largely unknown. In the present work we systematically investigate the impact of clathrin knockdown on CXCR4 internalization, signaling, and receptor post-translational modification. Inhibition of clathrin-mediated endocytosis (CME) significantly reduced CXCR4 internalization. In contrast to other receptors, clathrin knockdown increased CXCL12-dependent ERK1/2 signaling. Simultaneous inhibition of CME and lipid raft disruption abrogated this increase in ERK1/2 phosphorylation suggesting that endocytic pathway compensation can influence signaling outcomes. Interestingly, using an antibody sensitive to CXCR4 post-translational modification, we also found that our ability to detect CXCR4 was drastically reduced upon clathrin knockdown. We hypothesize that this effect was due to differences in receptor post-translational modification as total CXCR4 protein and mRNA levels were unchanged. Lastly, we show that clathrin knockdown reduced CXCL12-dependent cell migration irrespective of an observed increase in ERK1/2 phosphorylation. Altogether, this work supports a complex model by which modulation of endocytosis affects not only receptor signaling and internalization but also receptor post-translational modification.https://www.frontiersin.org/article/10.3389/fcell.2019.00077/fullCXCR4ERK signalingmembrane traffickingclathrinclathrin-mediated endocytosisG proteincoupled receptor |
| spellingShingle | Maxwell S. DeNies Luciana K. Rosselli-Murai Santiago Schnell Santiago Schnell Santiago Schnell Allen P. Liu Allen P. Liu Allen P. Liu Allen P. Liu Clathrin Heavy Chain Knockdown Impacts CXCR4 Signaling and Post-translational Modification Frontiers in Cell and Developmental Biology CXCR4 ERK signaling membrane trafficking clathrin clathrin-mediated endocytosis G proteincoupled receptor |
| title | Clathrin Heavy Chain Knockdown Impacts CXCR4 Signaling and Post-translational Modification |
| title_full | Clathrin Heavy Chain Knockdown Impacts CXCR4 Signaling and Post-translational Modification |
| title_fullStr | Clathrin Heavy Chain Knockdown Impacts CXCR4 Signaling and Post-translational Modification |
| title_full_unstemmed | Clathrin Heavy Chain Knockdown Impacts CXCR4 Signaling and Post-translational Modification |
| title_short | Clathrin Heavy Chain Knockdown Impacts CXCR4 Signaling and Post-translational Modification |
| title_sort | clathrin heavy chain knockdown impacts cxcr4 signaling and post translational modification |
| topic | CXCR4 ERK signaling membrane trafficking clathrin clathrin-mediated endocytosis G proteincoupled receptor |
| url | https://www.frontiersin.org/article/10.3389/fcell.2019.00077/full |
| work_keys_str_mv | AT maxwellsdenies clathrinheavychainknockdownimpactscxcr4signalingandposttranslationalmodification AT lucianakrossellimurai clathrinheavychainknockdownimpactscxcr4signalingandposttranslationalmodification AT santiagoschnell clathrinheavychainknockdownimpactscxcr4signalingandposttranslationalmodification AT santiagoschnell clathrinheavychainknockdownimpactscxcr4signalingandposttranslationalmodification AT santiagoschnell clathrinheavychainknockdownimpactscxcr4signalingandposttranslationalmodification AT allenpliu clathrinheavychainknockdownimpactscxcr4signalingandposttranslationalmodification AT allenpliu clathrinheavychainknockdownimpactscxcr4signalingandposttranslationalmodification AT allenpliu clathrinheavychainknockdownimpactscxcr4signalingandposttranslationalmodification AT allenpliu clathrinheavychainknockdownimpactscxcr4signalingandposttranslationalmodification |