Structural Organization and Dynamics of Homodimeric Cytohesin Family Arf GTPase Exchange Factors in Solution and on Membranes

Das, Sanchaita; Malaby, Andrew W.; Nawrotek, Agata; Zhang, Wenhua; Zeghouf, Mahel; Maslen, Sarah; Skehel, Mark; Chakravarthy, Srinivas; Irving, Thomas C.; Bilsel, Osman; Cherfils, Jacqueline; Lambright, David G.

doi:10.1016/j.str.2019.09.007

Title: Structural Organization and Dynamics of Homodimeric Cytohesin Family Arf GTPase Exchange Factors in Solution and on Membranes

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Abstract

Membrane dynamic processes require Arf GTPase activation by guanine nucleotide exchange factors (GEFs) with a Sec7 domain. Cytohesin family Arf GEFs function in signaling and cell migration through Arf GTPase activation on the plasma membrane and endosomes. In this study, the structural organization of two cytohesins (Grp1 and ARNO) was investigated in solution by size exclusion-small angle X-ray scattering and negative stain-electron microscopy and on membranes by dynamic light scattering, hydrogen-deuterium exchange-mass spectrometry and guanosine diphosphate (GDP)/guanosine triphosphate (GTP) exchange assays. The results suggest that cytohesins form elongated dimers with a central coiled coil and membrane-binding pleckstrin-homology (PH) domains at opposite ends. Here, the dimers display significant conformational heterogeneity, with a preference for compact to intermediate conformations. Phosphoinositide-dependent membrane recruitment is mediated by one PH domain at a time and alters the conformational dynamics to prime allosteric activation by Arf-GTP. A structural model for membrane targeting and allosteric activation of full-length cytohesin dimers is discussed.

Authors:: Das, Sanchaita; Malaby, Andrew W.; Nawrotek, Agata; Zhang, Wenhua; Zeghouf, Mahel; Maslen, Sarah; Skehel, Mark; Chakravarthy, Srinivas; Irving, Thomas C.; Bilsel, Osman; Cherfils, Jacqueline; Lambright, David G.

Publication Date:: Sun Dec 01 00:00:00 EST 2019

Research Org.:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Org.:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division; National Institute Health (NIH)

OSTI Identifier:: 1580152

Alternate Identifier(s):: OSTI ID: 1576873; OSTI ID: 1578202

Grant/Contract Number:: AC02-06CH11357; GM056324

Resource Type:: Published Article

Journal Name:: Structure

Additional Journal Information:: Journal Name: Structure Journal Volume: 27 Journal Issue: 12; Journal ID: ISSN 0969-2126

Publisher:: Elsevier

Country of Publication:: United Kingdom

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; GEF; SEC; SAXS; NS; EM; DLS; HDX; MS; autoinhibition; structure

Citation Formats


                    Das, Sanchaita, Malaby, Andrew W., Nawrotek, Agata, Zhang, Wenhua, Zeghouf, Mahel, Maslen, Sarah, Skehel, Mark, Chakravarthy, Srinivas, Irving, Thomas C., Bilsel, Osman, Cherfils, Jacqueline, and Lambright, David G. Structural Organization and Dynamics of Homodimeric Cytohesin Family Arf GTPase Exchange Factors in Solution and on Membranes.  United Kingdom: N. p., 2019. 
Web.  doi:10.1016/j.str.2019.09.007.

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                    Das, Sanchaita, Malaby, Andrew W., Nawrotek, Agata, Zhang, Wenhua, Zeghouf, Mahel, Maslen, Sarah, Skehel, Mark, Chakravarthy, Srinivas, Irving, Thomas C., Bilsel, Osman, Cherfils, Jacqueline, & Lambright, David G. Structural Organization and Dynamics of Homodimeric Cytohesin Family Arf GTPase Exchange Factors in Solution and on Membranes.  United Kingdom.  https://doi.org/10.1016/j.str.2019.09.007

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                    Das, Sanchaita, Malaby, Andrew W., Nawrotek, Agata, Zhang, Wenhua, Zeghouf, Mahel, Maslen, Sarah, Skehel, Mark, Chakravarthy, Srinivas, Irving, Thomas C., Bilsel, Osman, Cherfils, Jacqueline, and Lambright, David G. Sun .  
"Structural Organization and Dynamics of Homodimeric Cytohesin Family Arf GTPase Exchange Factors in Solution and on Membranes".  United Kingdom.  https://doi.org/10.1016/j.str.2019.09.007.

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@article{osti_1580152,

  title        = {Structural Organization and Dynamics of Homodimeric Cytohesin Family Arf GTPase Exchange Factors in Solution and on Membranes},

  author       = {Das, Sanchaita and Malaby, Andrew W. and Nawrotek, Agata and Zhang, Wenhua and Zeghouf, Mahel and Maslen, Sarah and Skehel, Mark and Chakravarthy, Srinivas and Irving, Thomas C. and Bilsel, Osman and Cherfils, Jacqueline and Lambright, David G.},

  abstractNote = {Membrane dynamic processes require Arf GTPase activation by guanine nucleotide exchange factors (GEFs) with a Sec7 domain. Cytohesin family Arf GEFs function in signaling and cell migration through Arf GTPase activation on the plasma membrane and endosomes. In this study, the structural organization of two cytohesins (Grp1 and ARNO) was investigated in solution by size exclusion-small angle X-ray scattering and negative stain-electron microscopy and on membranes by dynamic light scattering, hydrogen-deuterium exchange-mass spectrometry and guanosine diphosphate (GDP)/guanosine triphosphate (GTP) exchange assays. The results suggest that cytohesins form elongated dimers with a central coiled coil and membrane-binding pleckstrin-homology (PH) domains at opposite ends. Here, the dimers display significant conformational heterogeneity, with a preference for compact to intermediate conformations. Phosphoinositide-dependent membrane recruitment is mediated by one PH domain at a time and alters the conformational dynamics to prime allosteric activation by Arf-GTP. A structural model for membrane targeting and allosteric activation of full-length cytohesin dimers is discussed.},

  doi          = {10.1016/j.str.2019.09.007},

  journal      = {Structure},

  number       = 12,

  volume       = 27,

  place        = {United Kingdom},

  year         = {Sun Dec 01 00:00:00 EST 2019},

  month        = {Sun Dec 01 00:00:00 EST 2019}

}

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