Adaptation of the protein translational apparatus during ATDC5 chondrogenic differentiation
Introduction: Ribosome biogenesis is integrated with many cellular processes including proliferation, differentiation and oncogenic events. Chondrogenic proliferation and differentiation require a high cellular translational capacity to facilitate cartilaginous extracellular matrix production. We he...
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| Format: | Article |
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KeAi Communications Co., Ltd.
2022-06-01
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| Series: | Non-coding RNA Research |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2468054022000087 |
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| author | Mandy M.F. Steinbusch Guus G.H. van den Akker Andy Cremers Adhiambo M.A. Witlox Heleen M. Staal Mandy J. Peffers Lodewijk W. van Rhijn Marjolein M.J. Caron Tim J.M. Welting |
| author_facet | Mandy M.F. Steinbusch Guus G.H. van den Akker Andy Cremers Adhiambo M.A. Witlox Heleen M. Staal Mandy J. Peffers Lodewijk W. van Rhijn Marjolein M.J. Caron Tim J.M. Welting |
| author_sort | Mandy M.F. Steinbusch |
| collection | DOAJ |
| description | Introduction: Ribosome biogenesis is integrated with many cellular processes including proliferation, differentiation and oncogenic events. Chondrogenic proliferation and differentiation require a high cellular translational capacity to facilitate cartilaginous extracellular matrix production. We here investigated the expression dynamics of factors involved in ribosome biogenesis during in vitro chondrogenic differentiation and determined whether protein translation capacity adapts to different phases of chondrogenic differentiation. Materials: SnoRNA expression during ATDC5 differentiation was analyzed by RNA sequencing of samples acquired from day 0 (progenitor stage), 7 (chondrogenic stage) and day 14 (hypertrophic stage). RT-qPCR was used to determine expression of fibrillarin, dyskerin, UBF-1, Sox9, Col2a1, Runx2, Col10a1 mRNAs and 18S, 5.8S and 28S rRNAs. Protein expression of fibrillarin, dyskerin and UBF-1 was determined by immunoblotting. Ribosomal RNA content per cell was determined by calculating rRNA RT-qPCR signals relative to DNA content (SYBR Green assay). Total protein translational activity was evaluated with a puromycilation assay and polysome profiling. Results: As a result of initiation of chondrogenic differentiation (Δt0-t7), 21 snoRNAs were differentially expressed (DE). Hypertrophic differentiation caused DE of 23 snoRNAs (Δt7-t14) and 43 when t0 was compared to t14. DE snoRNAs, amongst others, target nucleotide modifications in the 28S rRNA peptidyl transferase center and the 18S rRNA decoding center. UBF-1, fibrillarin and dyskerin expression increased as function of differentiation and displayed highest fold induction at day 5–6 in differentiation. Ribosomal RNA content per cell was significantly increased at day 7, but not at day 14 in differentiation. Similar dynamics in translational capacity and monosomal ribosome fraction were observed during differentiation. Conclusion: The expression of a great number of ribosome biogenesis factors is altered during chondrogenic differentiation of ATDC5 cells, which is accompanied by significant changes in cellular translational activity. This elucidation of ribosome biogenesis dynamics in chondrogenic differentiation models enables the further understanding of the role of ribosome biogenesis and activity during chondrocyte cell commitment and their roles in human skeletal development diseases. |
| first_indexed | 2024-04-12T14:26:40Z |
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| institution | Directory Open Access Journal |
| issn | 2468-0540 |
| language | English |
| last_indexed | 2024-04-24T08:34:52Z |
| publishDate | 2022-06-01 |
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| spelling | doaj.art-bb29e52f629949dba5afb57af8b7828c2024-04-16T18:08:19ZengKeAi Communications Co., Ltd.Non-coding RNA Research2468-05402022-06-01725565Adaptation of the protein translational apparatus during ATDC5 chondrogenic differentiationMandy M.F. Steinbusch0Guus G.H. van den Akker1Andy Cremers2Adhiambo M.A. Witlox3Heleen M. Staal4Mandy J. Peffers5Lodewijk W. van Rhijn6Marjolein M.J. Caron7Tim J.M. Welting8Laboratory for Experimental Orthopedics, Department of Orthopedic Surgery, Maastricht University, P.O. Box 5800, 6202 AZ, Maastricht, the NetherlandsLaboratory for Experimental Orthopedics, Department of Orthopedic Surgery, Maastricht University, P.O. Box 5800, 6202 AZ, Maastricht, the NetherlandsLaboratory for Experimental Orthopedics, Department of Orthopedic Surgery, Maastricht University, P.O. Box 5800, 6202 AZ, Maastricht, the NetherlandsLaboratory for Experimental Orthopedics, Department of Orthopedic Surgery, Maastricht University Medical Center, P.O. Box 5800, 6202 AZ, Maastricht, the NetherlandsLaboratory for Experimental Orthopedics, Department of Orthopedic Surgery, Maastricht University Medical Center, P.O. Box 5800, 6202 AZ, Maastricht, the NetherlandsDepartment of Musculoskeletal Biology, Institute of Life Course and Medical Sciences, University of Liverpool, William Henry Duncan Building, 6 West Derby Street, L7 8TX, Liverpool, United KingdomLaboratory for Experimental Orthopedics, Department of Orthopedic Surgery, Maastricht University Medical Center, P.O. Box 5800, 6202 AZ, Maastricht, the NetherlandsLaboratory for Experimental Orthopedics, Department of Orthopedic Surgery, Maastricht University, P.O. Box 5800, 6202 AZ, Maastricht, the Netherlands; Corresponding author.Laboratory for Experimental Orthopedics, Department of Orthopedic Surgery, Maastricht University, P.O. Box 5800, 6202 AZ, Maastricht, the Netherlands; Laboratory for Experimental Orthopedics, Department of Orthopedic Surgery, Maastricht University Medical Center, P.O. Box 5800, 6202 AZ, Maastricht, the NetherlandsIntroduction: Ribosome biogenesis is integrated with many cellular processes including proliferation, differentiation and oncogenic events. Chondrogenic proliferation and differentiation require a high cellular translational capacity to facilitate cartilaginous extracellular matrix production. We here investigated the expression dynamics of factors involved in ribosome biogenesis during in vitro chondrogenic differentiation and determined whether protein translation capacity adapts to different phases of chondrogenic differentiation. Materials: SnoRNA expression during ATDC5 differentiation was analyzed by RNA sequencing of samples acquired from day 0 (progenitor stage), 7 (chondrogenic stage) and day 14 (hypertrophic stage). RT-qPCR was used to determine expression of fibrillarin, dyskerin, UBF-1, Sox9, Col2a1, Runx2, Col10a1 mRNAs and 18S, 5.8S and 28S rRNAs. Protein expression of fibrillarin, dyskerin and UBF-1 was determined by immunoblotting. Ribosomal RNA content per cell was determined by calculating rRNA RT-qPCR signals relative to DNA content (SYBR Green assay). Total protein translational activity was evaluated with a puromycilation assay and polysome profiling. Results: As a result of initiation of chondrogenic differentiation (Δt0-t7), 21 snoRNAs were differentially expressed (DE). Hypertrophic differentiation caused DE of 23 snoRNAs (Δt7-t14) and 43 when t0 was compared to t14. DE snoRNAs, amongst others, target nucleotide modifications in the 28S rRNA peptidyl transferase center and the 18S rRNA decoding center. UBF-1, fibrillarin and dyskerin expression increased as function of differentiation and displayed highest fold induction at day 5–6 in differentiation. Ribosomal RNA content per cell was significantly increased at day 7, but not at day 14 in differentiation. Similar dynamics in translational capacity and monosomal ribosome fraction were observed during differentiation. Conclusion: The expression of a great number of ribosome biogenesis factors is altered during chondrogenic differentiation of ATDC5 cells, which is accompanied by significant changes in cellular translational activity. This elucidation of ribosome biogenesis dynamics in chondrogenic differentiation models enables the further understanding of the role of ribosome biogenesis and activity during chondrocyte cell commitment and their roles in human skeletal development diseases.http://www.sciencedirect.com/science/article/pii/S2468054022000087ATDC5Chondrogenic differentiationRibosomeTranslationsnoRNA |
| spellingShingle | Mandy M.F. Steinbusch Guus G.H. van den Akker Andy Cremers Adhiambo M.A. Witlox Heleen M. Staal Mandy J. Peffers Lodewijk W. van Rhijn Marjolein M.J. Caron Tim J.M. Welting Adaptation of the protein translational apparatus during ATDC5 chondrogenic differentiation Non-coding RNA Research ATDC5 Chondrogenic differentiation Ribosome Translation snoRNA |
| title | Adaptation of the protein translational apparatus during ATDC5 chondrogenic differentiation |
| title_full | Adaptation of the protein translational apparatus during ATDC5 chondrogenic differentiation |
| title_fullStr | Adaptation of the protein translational apparatus during ATDC5 chondrogenic differentiation |
| title_full_unstemmed | Adaptation of the protein translational apparatus during ATDC5 chondrogenic differentiation |
| title_short | Adaptation of the protein translational apparatus during ATDC5 chondrogenic differentiation |
| title_sort | adaptation of the protein translational apparatus during atdc5 chondrogenic differentiation |
| topic | ATDC5 Chondrogenic differentiation Ribosome Translation snoRNA |
| url | http://www.sciencedirect.com/science/article/pii/S2468054022000087 |
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