Actinin BioID reveals sarcomere crosstalk with oxidative metabolism through interactions with IGF2BP2
Actinins are strain-sensing actin cross-linkers that are ubiquitously expressed and harbor mutations in human diseases. We utilize CRISPR, pluripotent stem cells, and BioID to study actinin interactomes in human cardiomyocytes. We identify 324 actinin proximity partners, including those that are dep...
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Published in | Cell reports (Cambridge) Vol. 36; no. 6; p. 109512 |
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Main Authors | , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Elsevier Inc
10.08.2021
Elsevier |
Subjects | |
Online Access | Get full text |
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Summary: | Actinins are strain-sensing actin cross-linkers that are ubiquitously expressed and harbor mutations in human diseases. We utilize CRISPR, pluripotent stem cells, and BioID to study actinin interactomes in human cardiomyocytes. We identify 324 actinin proximity partners, including those that are dependent on sarcomere assembly. We confirm 19 known interactors and identify a network of RNA-binding proteins, including those with RNA localization functions. In vivo and biochemical interaction studies support that IGF2BP2 localizes electron transport chain transcripts to actinin neighborhoods through interactions between its K homology (KH) domain and actinin’s rod domain. We combine alanine scanning mutagenesis and metabolic assays to disrupt and functionally interrogate actinin-IGF2BP2 interactions, which reveal an essential role in metabolic responses to pathological sarcomere activation using a hypertrophic cardiomyopathy model. This study expands our functional knowledge of actinin, uncovers sarcomere interaction partners, and reveals sarcomere crosstalk with IGF2BP2 for metabolic adaptation relevant to human disease.
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•BioID identifies 324 actinin proximity partners through sarcomere assembly•IGF2BP2 directly binds actinin’s rod domain through its KH domain•IGF2BP2-actinin interactions determine ETC transcript localization•Actinin-IGF2BP2 interactions regulate metabolic adaptation to sarcomere function
Ladha et al. combine BioID, human cardiomyocytes, and CRISPR-Cas9 to interrogate the actinin interactome. This reveals 324 actinin proximity partners, including RNA-binding proteins that bind transcripts encoding ETC functional components. Among these RNA-binding proteins, IGF2BP2 directly binds actinin, and actinin-IGF2BP2 interactions regulate ETC transcript localization and metabolic adaptation to sarcomere function. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 AUTHOR CONTRIBUTIONS Investigation and validation, F.A.L., A.M.P., K.T., N.L., and J.T.H.; cells and/or reagent generation, F.A.L., A.M.P., K.T., N.L., S.G., R.C., R.R., E.M., Y.S.C., and J.T.H.; formal analysis, F.A.L., A.M.P., and J.T.H.; funding acquisition, F.A.L., A.M.P., and J.T.H.; conceptualization and supervision, J.T.H. and F.A.L.; writing, F.A.L., A.M.P., and J.T.H.; and all authors reviewed the manuscript prior to submission. |
ISSN: | 2211-1247 2211-1247 |
DOI: | 10.1016/j.celrep.2021.109512 |