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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...

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Autores principales: Ferdous, Ahlan S., Costa Dos Santos, Stephany J., Kanzler, Charlotte R., Shin, Heaji, Carrick, Brian H., Crittenden, Sarah L., Wickens, Marvin, Kimble, Judith
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
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.
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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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