The CXCR4-Dependent LASP1-Ago2 Interaction in Triple-Negative Breast Cancer
The CXCR4-LASP1 axis is an emerging target in the field of breast cancer metastasis. C-X-C chemokine receptor type 4 (CXCR4) mediates directed cell migration when activated by its cognate ligand CXCL12. LIM and SH3 Protein 1 (LASP1) is a critical node in the CXCR4 signaling pathway, as its deficienc...
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| Format: | Article |
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MDPI AG
2020-08-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/12/9/2455 |
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| author | Augustus M. C. Tilley Cory M. Howard Sangita Sridharan Boopathi Subramaniyan Nicole R. Bearss Sawsan Alkhalili Dayanidhi Raman |
| author_facet | Augustus M. C. Tilley Cory M. Howard Sangita Sridharan Boopathi Subramaniyan Nicole R. Bearss Sawsan Alkhalili Dayanidhi Raman |
| author_sort | Augustus M. C. Tilley |
| collection | DOAJ |
| description | The CXCR4-LASP1 axis is an emerging target in the field of breast cancer metastasis. C-X-C chemokine receptor type 4 (CXCR4) mediates directed cell migration when activated by its cognate ligand CXCL12. LIM and SH3 Protein 1 (LASP1) is a critical node in the CXCR4 signaling pathway, as its deficiency blocks CXCR4-dependent Matrigel invasion. The mechanism by which LASP1 facilitates this invasive ability of tumor cells when CXCR4 is activated is unknown. Our previous proteomics work had revealed several components of the RNA interference (RNAi) machinery as being potential LASP1 interacting proteins. Here we report that argonaute 2 (Ago2), a protein with central involvement in RNAi, associates with LASP1 in triple-negative breast cancer (TNBC) cells. We demonstrate that LASP1 co-immunoprecipitates with Ago2 endogenously in a CXCL12-dependent manner, with further confirmation of this interaction by proximity ligation assay. Furthermore, this association is specific to CXCR4 as it can be abrogated by the CXCR4 antagonist, AMD3465. By GST-pulldown approach, we identify that LASP1 directly binds to Ago2 through its LIM and SH3 domains, and that this binding is dictated by the S146 and Y171 phosphorylation sites of LASP1. Additionally, the phosphorylation status of LASP1 affected tumor suppressor microRNA (miRNA) Let-7a-guided Ago2 activity. Levels of several endogenous targets of Let-7a were found to be altered including C-C chemokine receptor type 7 (CCR7), which is another critical chemokine receptor involved in metastasis to lymph nodes. Our results suggest a novel role for the LASP1-Ago2 module in shaping the RNAi landscape, functionally impacting the invasive ability of cancer cells. |
| first_indexed | 2024-03-10T16:42:58Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-10T16:42:58Z |
| publishDate | 2020-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-7ea16dd3246946b0ba950de422ca9bef2023-11-20T11:50:27ZengMDPI AGCancers2072-66942020-08-01129245510.3390/cancers12092455The CXCR4-Dependent LASP1-Ago2 Interaction in Triple-Negative Breast CancerAugustus M. C. Tilley0Cory M. Howard1Sangita Sridharan2Boopathi Subramaniyan3Nicole R. Bearss4Sawsan Alkhalili5Dayanidhi Raman6Department of Cancer Biology, College of Medicine and Life Sciences, University of Toledo Health Science Campus, Toledo, OH 43614, USADepartment of Cancer Biology, College of Medicine and Life Sciences, University of Toledo Health Science Campus, Toledo, OH 43614, USADepartment of Cancer Biology, College of Medicine and Life Sciences, University of Toledo Health Science Campus, Toledo, OH 43614, USADepartment of Cancer Biology, College of Medicine and Life Sciences, University of Toledo Health Science Campus, Toledo, OH 43614, USADepartment of Cancer Biology, College of Medicine and Life Sciences, University of Toledo Health Science Campus, Toledo, OH 43614, USADepartment of Cancer Biology, College of Medicine and Life Sciences, University of Toledo Health Science Campus, Toledo, OH 43614, USADepartment of Cancer Biology, College of Medicine and Life Sciences, University of Toledo Health Science Campus, Toledo, OH 43614, USAThe CXCR4-LASP1 axis is an emerging target in the field of breast cancer metastasis. C-X-C chemokine receptor type 4 (CXCR4) mediates directed cell migration when activated by its cognate ligand CXCL12. LIM and SH3 Protein 1 (LASP1) is a critical node in the CXCR4 signaling pathway, as its deficiency blocks CXCR4-dependent Matrigel invasion. The mechanism by which LASP1 facilitates this invasive ability of tumor cells when CXCR4 is activated is unknown. Our previous proteomics work had revealed several components of the RNA interference (RNAi) machinery as being potential LASP1 interacting proteins. Here we report that argonaute 2 (Ago2), a protein with central involvement in RNAi, associates with LASP1 in triple-negative breast cancer (TNBC) cells. We demonstrate that LASP1 co-immunoprecipitates with Ago2 endogenously in a CXCL12-dependent manner, with further confirmation of this interaction by proximity ligation assay. Furthermore, this association is specific to CXCR4 as it can be abrogated by the CXCR4 antagonist, AMD3465. By GST-pulldown approach, we identify that LASP1 directly binds to Ago2 through its LIM and SH3 domains, and that this binding is dictated by the S146 and Y171 phosphorylation sites of LASP1. Additionally, the phosphorylation status of LASP1 affected tumor suppressor microRNA (miRNA) Let-7a-guided Ago2 activity. Levels of several endogenous targets of Let-7a were found to be altered including C-C chemokine receptor type 7 (CCR7), which is another critical chemokine receptor involved in metastasis to lymph nodes. Our results suggest a novel role for the LASP1-Ago2 module in shaping the RNAi landscape, functionally impacting the invasive ability of cancer cells.https://www.mdpi.com/2072-6694/12/9/2455Argonaute2LASP1CXCR4Let-7atriple negative breast cancer |
| spellingShingle | Augustus M. C. Tilley Cory M. Howard Sangita Sridharan Boopathi Subramaniyan Nicole R. Bearss Sawsan Alkhalili Dayanidhi Raman The CXCR4-Dependent LASP1-Ago2 Interaction in Triple-Negative Breast Cancer Cancers Argonaute2 LASP1 CXCR4 Let-7a triple negative breast cancer |
| title | The CXCR4-Dependent LASP1-Ago2 Interaction in Triple-Negative Breast Cancer |
| title_full | The CXCR4-Dependent LASP1-Ago2 Interaction in Triple-Negative Breast Cancer |
| title_fullStr | The CXCR4-Dependent LASP1-Ago2 Interaction in Triple-Negative Breast Cancer |
| title_full_unstemmed | The CXCR4-Dependent LASP1-Ago2 Interaction in Triple-Negative Breast Cancer |
| title_short | The CXCR4-Dependent LASP1-Ago2 Interaction in Triple-Negative Breast Cancer |
| title_sort | cxcr4 dependent lasp1 ago2 interaction in triple negative breast cancer |
| topic | Argonaute2 LASP1 CXCR4 Let-7a triple negative breast cancer |
| url | https://www.mdpi.com/2072-6694/12/9/2455 |
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