期刊
ELIFE
卷 11, 期 -, 页码 -出版社
eLIFE SCIENCES PUBL LTD
DOI: 10.7554/eLife.80217
关键词
dynein; p150 Glued; multivalency; protein disorder; NMR; protein interactions
类别
资金
- National Science Foundation [1617019, 2003557]
- National Institute of Biological Resources [1S10OD018518]
- Division Of Chemistry
- Direct For Mathematical & Physical Scien [2003557] Funding Source: National Science Foundation
- Div Of Molecular and Cellular Bioscience
- Direct For Biological Sciences [1617019] Funding Source: National Science Foundation
Cytoplasmic dynein is crucial for intracellular transport, with its intermediate chain (IC) regulating interactions with p150(Glued) and NudE. The multivalent binding of light chains modulates autoinhibition, highlighting their importance in IC assembly and organization.
As the only major retrograde transporter along microtubules, cytoplasmic dynein plays crucial roles in the intracellular transport of organelles and other cargoes. Central to the function of this motor protein complex is dynein intermediate chain (IC), which binds the three dimeric dynein light chains at multivalent sites, and dynactin p150(Glued) and nuclear distribution protein (NudE) at overlapping sites of its intrinsically disordered N-terminal domain. The disorder in IC has hindered cryo-electron microscopy and X-ray crystallography studies of its structure and interactions. Here we use a suite of biophysical methods to reveal how multivalent binding of the three light chains regulates IC interactions with p150(Glued) and NudE. Using IC from Chaetomium thermophilum, a tractable species to interrogate IC interactions, we identify a significant reduction in binding affinity of IC to p150(Glued) and a loss of binding to NudE for constructs containing the entire N-terminal domain as well as for full-length constructs when compared to the tight binding observed with short IC constructs. We attribute this difference to autoinhibition caused by long-range intramolecular interactions between the N-terminal single alpha-helix of IC, the common site for p150(Glued), and NudE binding, and residues closer to the end of the N-terminal domain. Reconstitution of IC subcomplexes demonstrates that autoinhibition is differentially regulated by light chains binding, underscoring their importance both in assembly and organization of IC, and in selection between multiple binding partners at the same site.
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