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Functional significance of PUF partnerships in C. elegans germline stem cells
PUF RNA-binding proteins are conserved stem cell regulators. Four PUF proteins govern self-renewal of C. elegans germline stem cells together with two intrinsically disordered proteins, LST-1 and SYGL-1. Based on yeast two-hybrid results, we proposed a composite self-renewal hub in the stem cell reg...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9949348/ https://www.ncbi.nlm.nih.gov/pubmed/36824876 http://dx.doi.org/10.1101/2023.02.15.528708 |
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author | Ferdous, Ahlan S. Costa Dos Santos, Stephany J. Kanzler, Charlotte R. Shin, Heaji Carrick, Brian H. Crittenden, Sarah L. Wickens, Marvin Kimble, Judith |
author_facet | Ferdous, Ahlan S. Costa Dos Santos, Stephany J. Kanzler, Charlotte R. Shin, Heaji Carrick, Brian H. Crittenden, Sarah L. Wickens, Marvin Kimble, Judith |
author_sort | Ferdous, Ahlan S. |
collection | PubMed |
description | PUF RNA-binding proteins are conserved stem cell regulators. Four PUF proteins govern self-renewal of C. elegans germline stem cells together with two intrinsically disordered proteins, LST-1 and SYGL-1. Based on yeast two-hybrid results, we proposed a composite self-renewal hub in the stem cell regulatory network, with eight PUF partnerships and extensive redundancy. Here, we investigate LST-1–PUF and SYGL-1–PUF partnerships and their molecular activities in their natural context – nematode stem cells. We confirm LST-1–PUF partnerships and their specificity to self-renewal PUFs by co-immunoprecipitation and show that an LST-1(A(m)B(m)) mutant defective for PUF-interacting motifs does not complex with PUFs in nematodes. LST-1(A(m)B(m)) is used to explore the functional significance of the LST-1–PUF partnership. Tethered LST-1 requires the partnership to repress expression of a reporter RNA, and LST-1 requires the partnership to co-immunoprecipitate with NTL-1/Not1 of the CCR4-NOT complex. We suggest that the partnership provides multiple molecular interactions that work together to form an effector complex on PUF target RNAs. Comparison of PUF-LST-1 and Pumilio–Nanos reveals fundamental molecular differences, making PUF–LST-1 a distinct paradigm for PUF partnerships. |
format | Online Article Text |
id | pubmed-9949348 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Cold Spring Harbor Laboratory |
record_format | MEDLINE/PubMed |
spelling | pubmed-99493482023-02-24 Functional significance of PUF partnerships in C. elegans germline stem cells Ferdous, Ahlan S. Costa Dos Santos, Stephany J. Kanzler, Charlotte R. Shin, Heaji Carrick, Brian H. Crittenden, Sarah L. Wickens, Marvin Kimble, Judith bioRxiv Article PUF RNA-binding proteins are conserved stem cell regulators. Four PUF proteins govern self-renewal of C. elegans germline stem cells together with two intrinsically disordered proteins, LST-1 and SYGL-1. Based on yeast two-hybrid results, we proposed a composite self-renewal hub in the stem cell regulatory network, with eight PUF partnerships and extensive redundancy. Here, we investigate LST-1–PUF and SYGL-1–PUF partnerships and their molecular activities in their natural context – nematode stem cells. We confirm LST-1–PUF partnerships and their specificity to self-renewal PUFs by co-immunoprecipitation and show that an LST-1(A(m)B(m)) mutant defective for PUF-interacting motifs does not complex with PUFs in nematodes. LST-1(A(m)B(m)) is used to explore the functional significance of the LST-1–PUF partnership. Tethered LST-1 requires the partnership to repress expression of a reporter RNA, and LST-1 requires the partnership to co-immunoprecipitate with NTL-1/Not1 of the CCR4-NOT complex. We suggest that the partnership provides multiple molecular interactions that work together to form an effector complex on PUF target RNAs. Comparison of PUF-LST-1 and Pumilio–Nanos reveals fundamental molecular differences, making PUF–LST-1 a distinct paradigm for PUF partnerships. Cold Spring Harbor Laboratory 2023-02-16 /pmc/articles/PMC9949348/ /pubmed/36824876 http://dx.doi.org/10.1101/2023.02.15.528708 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator. |
spellingShingle | Article Ferdous, Ahlan S. Costa Dos Santos, Stephany J. Kanzler, Charlotte R. Shin, Heaji Carrick, Brian H. Crittenden, Sarah L. Wickens, Marvin Kimble, Judith Functional significance of PUF partnerships in C. elegans germline stem cells |
title | Functional significance of PUF partnerships in C. elegans germline stem cells |
title_full | Functional significance of PUF partnerships in C. elegans germline stem cells |
title_fullStr | Functional significance of PUF partnerships in C. elegans germline stem cells |
title_full_unstemmed | Functional significance of PUF partnerships in C. elegans germline stem cells |
title_short | Functional significance of PUF partnerships in C. elegans germline stem cells |
title_sort | functional significance of puf partnerships in c. elegans germline stem cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9949348/ https://www.ncbi.nlm.nih.gov/pubmed/36824876 http://dx.doi.org/10.1101/2023.02.15.528708 |
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