CacyBP/SIP - RPL6 interaction: potential influence on ribosome function

Abstract Previously, we have shown that CacyBP/SIP interacts with NPM1, a protein involved in ribosome biogenesis. In this work, we extended our previous studies to look for the potential impact of CacyBP/SIP on ribosome biogenesis and/or function. Using mass spectrometry analysis, we have found tha...

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Main Authors: Ewelina Jurewicz, Małgorzata Maksymowicz-Trivedi, Omid Saberi-Khomami, Olga Iwańska, Agata Starosta, Ewa Kilańczyk, Paweł Bieganowski, Adam Jarmuła, Wiesława Leśniak, Sławomir Filipek, Anna Filipek
Format: Article
Language:English
Published: Springer 2025-07-01
Series:Amino Acids
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Online Access:https://doi.org/10.1007/s00726-025-03464-3
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author Ewelina Jurewicz
Małgorzata Maksymowicz-Trivedi
Omid Saberi-Khomami
Olga Iwańska
Agata Starosta
Ewa Kilańczyk
Paweł Bieganowski
Adam Jarmuła
Wiesława Leśniak
Sławomir Filipek
Anna Filipek
author_facet Ewelina Jurewicz
Małgorzata Maksymowicz-Trivedi
Omid Saberi-Khomami
Olga Iwańska
Agata Starosta
Ewa Kilańczyk
Paweł Bieganowski
Adam Jarmuła
Wiesława Leśniak
Sławomir Filipek
Anna Filipek
author_sort Ewelina Jurewicz
collection DOAJ
description Abstract Previously, we have shown that CacyBP/SIP interacts with NPM1, a protein involved in ribosome biogenesis. In this work, we extended our previous studies to look for the potential impact of CacyBP/SIP on ribosome biogenesis and/or function. Using mass spectrometry analysis, we have found that several RPs could be potential CacyBP/SIP targets. Since RPL6 was one of the proteins with the best quality scores identified in this analysis we focused on the possible interaction between CacyBP/SIP and RPL6. By applying various biochemical methods, we confirmed this interaction and showed that it was direct. Moreover, in silico analysis allowed us to establish the domains/fragments of both proteins involved in the binding. To further explore the possible role of CacyBP/SIP in ribosome function we performed several analyses using neuroblastoma NB2a cell line with stably silenced CacyBP/SIP expression. We have found, by applying OPP (O-propargyl-puromycin), which labels nascent polypeptides, that the number of cells with enhanced staining in the perinuclear area, reminiscent of rough ER localization, was significantly lower in the cell line with diminished CacyBP/SIP level. To verify the influence of CacyBP/SIP on the efficiency of protein synthesis we investigated the level of Hsp70, a stress-inducible protein, in NB2a cells subjected to heat shock. The results, showing markedly higher Hsp70 production in control cells, indicate that CacyBP/SIP, most probably through interaction with RPL6 and/or other RPs, may have some influence on ribosome function and, possibly, on protein synthesis in the cell.
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spelling doaj-art-457f78bc4d9245b3a11d3aae4434ebba2025-08-20T03:05:14ZengSpringerAmino Acids1438-21992025-07-0157111810.1007/s00726-025-03464-3CacyBP/SIP - RPL6 interaction: potential influence on ribosome functionEwelina Jurewicz0Małgorzata Maksymowicz-Trivedi1Omid Saberi-Khomami2Olga Iwańska3Agata Starosta4Ewa Kilańczyk5Paweł Bieganowski6Adam Jarmuła7Wiesława Leśniak8Sławomir Filipek9Anna Filipek10Nencki Institute of Experimental Biology, Polish Academy of SciencesNencki Institute of Experimental Biology, Polish Academy of SciencesNencki Institute of Experimental Biology, Polish Academy of SciencesInstitute of Biochemistry and Biophysics, Polish Academy of SciencesInstitute of Biochemistry and Biophysics, Polish Academy of SciencesNencki Institute of Experimental Biology, Polish Academy of SciencesMossakowski Medical Research Institute, Polish Academy of SciencesNencki Institute of Experimental Biology, Polish Academy of SciencesNencki Institute of Experimental Biology, Polish Academy of SciencesFaculty of Chemistry, Biological and Chemical Research Centre, University of WarsawNencki Institute of Experimental Biology, Polish Academy of SciencesAbstract Previously, we have shown that CacyBP/SIP interacts with NPM1, a protein involved in ribosome biogenesis. In this work, we extended our previous studies to look for the potential impact of CacyBP/SIP on ribosome biogenesis and/or function. Using mass spectrometry analysis, we have found that several RPs could be potential CacyBP/SIP targets. Since RPL6 was one of the proteins with the best quality scores identified in this analysis we focused on the possible interaction between CacyBP/SIP and RPL6. By applying various biochemical methods, we confirmed this interaction and showed that it was direct. Moreover, in silico analysis allowed us to establish the domains/fragments of both proteins involved in the binding. To further explore the possible role of CacyBP/SIP in ribosome function we performed several analyses using neuroblastoma NB2a cell line with stably silenced CacyBP/SIP expression. We have found, by applying OPP (O-propargyl-puromycin), which labels nascent polypeptides, that the number of cells with enhanced staining in the perinuclear area, reminiscent of rough ER localization, was significantly lower in the cell line with diminished CacyBP/SIP level. To verify the influence of CacyBP/SIP on the efficiency of protein synthesis we investigated the level of Hsp70, a stress-inducible protein, in NB2a cells subjected to heat shock. The results, showing markedly higher Hsp70 production in control cells, indicate that CacyBP/SIP, most probably through interaction with RPL6 and/or other RPs, may have some influence on ribosome function and, possibly, on protein synthesis in the cell.https://doi.org/10.1007/s00726-025-03464-3CacyBP/SIPMolecular modelingProtein synthesisRibosomeRPL6
spellingShingle Ewelina Jurewicz
Małgorzata Maksymowicz-Trivedi
Omid Saberi-Khomami
Olga Iwańska
Agata Starosta
Ewa Kilańczyk
Paweł Bieganowski
Adam Jarmuła
Wiesława Leśniak
Sławomir Filipek
Anna Filipek
CacyBP/SIP - RPL6 interaction: potential influence on ribosome function
Amino Acids
CacyBP/SIP
Molecular modeling
Protein synthesis
Ribosome
RPL6
title CacyBP/SIP - RPL6 interaction: potential influence on ribosome function
title_full CacyBP/SIP - RPL6 interaction: potential influence on ribosome function
title_fullStr CacyBP/SIP - RPL6 interaction: potential influence on ribosome function
title_full_unstemmed CacyBP/SIP - RPL6 interaction: potential influence on ribosome function
title_short CacyBP/SIP - RPL6 interaction: potential influence on ribosome function
title_sort cacybp sip rpl6 interaction potential influence on ribosome function
topic CacyBP/SIP
Molecular modeling
Protein synthesis
Ribosome
RPL6
url https://doi.org/10.1007/s00726-025-03464-3
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