Phase separation of a PKA type I regulatory subunit regulates β-cell function through cAMP compartmentalization.

Cyclic adenosine monophosphate (cAMP), a ubiquitous second messenger, regulates a variety of cellular functions with high specificity. We previously showed that the type I regulatory subunit of cAMP-dependent protein kinase A (PKA), RIα, undergoes liquid-liquid phase separation (LLPS) to facilitate...

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Main Authors: Ha Neul Lee, Julia C Hardy, Emily H Pool, Jin-Fan Zhang, Su Hyun Kim, William F Buhl, Jessica G H Bruystens, Sohum Mehta, Susan S Taylor, Jin Zhang
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2025-07-01
Series:PLoS Biology
Online Access:https://doi.org/10.1371/journal.pbio.3003262
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author Ha Neul Lee
Julia C Hardy
Emily H Pool
Jin-Fan Zhang
Su Hyun Kim
William F Buhl
Jessica G H Bruystens
Sohum Mehta
Susan S Taylor
Jin Zhang
author_facet Ha Neul Lee
Julia C Hardy
Emily H Pool
Jin-Fan Zhang
Su Hyun Kim
William F Buhl
Jessica G H Bruystens
Sohum Mehta
Susan S Taylor
Jin Zhang
author_sort Ha Neul Lee
collection DOAJ
description Cyclic adenosine monophosphate (cAMP), a ubiquitous second messenger, regulates a variety of cellular functions with high specificity. We previously showed that the type I regulatory subunit of cAMP-dependent protein kinase A (PKA), RIα, undergoes liquid-liquid phase separation (LLPS) to facilitate spatial compartmentalization of cAMP. However, how RIα LLPS regulates cellular function is largely unknown. Here, we identify the formation of RIα condensates in MIN6 β cells and reveal key roles for RIα LLPS in regulating β cell function. By combining CRISPR-based RIα knockout with an RIα mutant (Y122A) that exhibits defective cAMP-induced LLPS, we demonstrate that RIα LLPS drives cAMP compartmentalization to tune β cell Ca2+ and cAMP oscillation frequency, control insulin secretion, regulate CREB-mediated gene expression and prevent uncontrolled proliferation. Our data establish the Y122A mutant as a selective molecular tool for studying RIα LLPS and expand our understanding of the functional impact of LLPS-driven protein assemblies.
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institution Kabale University
issn 1544-9173
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language English
publishDate 2025-07-01
publisher Public Library of Science (PLoS)
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spelling doaj-art-51bbf9d8224e4afea035cf2c0f1d0ec42025-08-20T03:57:59ZengPublic Library of Science (PLoS)PLoS Biology1544-91731545-78852025-07-01237e300326210.1371/journal.pbio.3003262Phase separation of a PKA type I regulatory subunit regulates β-cell function through cAMP compartmentalization.Ha Neul LeeJulia C HardyEmily H PoolJin-Fan ZhangSu Hyun KimWilliam F BuhlJessica G H BruystensSohum MehtaSusan S TaylorJin ZhangCyclic adenosine monophosphate (cAMP), a ubiquitous second messenger, regulates a variety of cellular functions with high specificity. We previously showed that the type I regulatory subunit of cAMP-dependent protein kinase A (PKA), RIα, undergoes liquid-liquid phase separation (LLPS) to facilitate spatial compartmentalization of cAMP. However, how RIα LLPS regulates cellular function is largely unknown. Here, we identify the formation of RIα condensates in MIN6 β cells and reveal key roles for RIα LLPS in regulating β cell function. By combining CRISPR-based RIα knockout with an RIα mutant (Y122A) that exhibits defective cAMP-induced LLPS, we demonstrate that RIα LLPS drives cAMP compartmentalization to tune β cell Ca2+ and cAMP oscillation frequency, control insulin secretion, regulate CREB-mediated gene expression and prevent uncontrolled proliferation. Our data establish the Y122A mutant as a selective molecular tool for studying RIα LLPS and expand our understanding of the functional impact of LLPS-driven protein assemblies.https://doi.org/10.1371/journal.pbio.3003262
spellingShingle Ha Neul Lee
Julia C Hardy
Emily H Pool
Jin-Fan Zhang
Su Hyun Kim
William F Buhl
Jessica G H Bruystens
Sohum Mehta
Susan S Taylor
Jin Zhang
Phase separation of a PKA type I regulatory subunit regulates β-cell function through cAMP compartmentalization.
PLoS Biology
title Phase separation of a PKA type I regulatory subunit regulates β-cell function through cAMP compartmentalization.
title_full Phase separation of a PKA type I regulatory subunit regulates β-cell function through cAMP compartmentalization.
title_fullStr Phase separation of a PKA type I regulatory subunit regulates β-cell function through cAMP compartmentalization.
title_full_unstemmed Phase separation of a PKA type I regulatory subunit regulates β-cell function through cAMP compartmentalization.
title_short Phase separation of a PKA type I regulatory subunit regulates β-cell function through cAMP compartmentalization.
title_sort phase separation of a pka type i regulatory subunit regulates β cell function through camp compartmentalization
url https://doi.org/10.1371/journal.pbio.3003262
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