Clathrin light chains are required for the gyrating-clathrin recycling pathway and thereby promote cell migration.
The clathrin light chain (CLC) subunits participate in several membrane traffic pathways involving both clathrin and actin, through binding the actin-organizing huntingtin-interacting proteins (Hip). However, CLCs are dispensable for clathrin-mediated endocytosis of many cargoes. Here we observe that CLC depletion affects cell migration through Hip binding and reduces surface expression of β1-integrin by interference with recycling following normal endocytosis of inactive β1-integrin. CLC depletion and expression of a modified CLC also inhibit the appearance of gyrating (G)-clathrin structures, known mediators of rapid recycling of transferrin receptor from endosomes. Expression of the modified CLC reduces β1-integrin and transferrin receptor recycling, as well as cell migration, implicating G-clathrin in these processes. Supporting a physiological role for CLC in migration, the CLCb isoform of CLC is upregulated in migratory human trophoblast cells during uterine invasion. Together, these studies establish CLCs as mediating clathrin-actin interactions needed for recycling by G-clathrin during migration.
1] Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California 94143, USA [2] Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California 94143, USA [3] Department of Microbiology and Immunology, University of California San Francisco, San Francisco, California 94143, USA [4] The G.W. Hooper Foundation, University of California San Francisco, San Francisco, California 94143, USA.
Sophia R
SR
Majeed
1] Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California 94143, USA [2] Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California 94143, USA [3] Department of Microbiology and Immunology, University of California San Francisco, San Francisco, California 94143, USA [4] The G.W. Hooper Foundation, University of California San Francisco, San Francisco, California 94143, USA [5].
Lavanya
L
Vasudevan
1] Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California 94143, USA [2] Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California 94143, USA [3] Department of Microbiology and Immunology, University of California San Francisco, San Francisco, California 94143, USA [4] The G.W. Hooper Foundation, University of California San Francisco, San Francisco, California 94143, USA [5].
Chih-Ying
CY
Chen
1] Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA [2].
Yi
Y
Luo
1] Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California 94143, USA [2] Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California 94143, USA [3] Department of Microbiology and Immunology, University of California San Francisco, San Francisco, California 94143, USA [4] The G.W. Hooper Foundation, University of California San Francisco, San Francisco, California 94143, USA.
Jorge A
JA
Torres
1] Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California 94143, USA [2] Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California 94143, USA [3] Department of Microbiology and Immunology, University of California San Francisco, San Francisco, California 94143, USA [4] The G.W. Hooper Foundation, University of California San Francisco, San Francisco, California 94143, USA.
Timothy M
TM
Evans
Department of Pathology and Centre for Trophoblast Research, University of Cambridge, Cambridge CB2 1QP, UK.
Andrew
A
Sharkey
1] Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California 94143, USA [2] Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California 94143, USA [3] Department of Microbiology and Immunology, University of California San Francisco, San Francisco, California 94143, USA [4] The G.W. Hooper Foundation, University of California San Francisco, San Francisco, California 94143, USA.
Amy B
AB
Foraker
1] Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California 94143, USA [2] Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California 94143, USA [3] Department of Microbiology and Immunology, University of California San Francisco, San Francisco, California 94143, USA [4] The G.W. Hooper Foundation, University of California San Francisco, San Francisco, California 94143, USA.
Nicole M L
NM
Wong
1] Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California 94143, USA [2] Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California 94143, USA [3] Department of Microbiology and Immunology, University of California San Francisco, San Francisco, California 94143, USA [4] The G.W. Hooper Foundation, University of California San Francisco, San Francisco, California 94143, USA.
Christopher
C
Esk
Department of Orthopedic Surgery, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Theresa A
TA
Freeman
Department of Pathology and Centre for Trophoblast Research, University of Cambridge, Cambridge CB2 1QP, UK.
Ashley
A
Moffett
Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
James H
JH
Keen
1] Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California 94143, USA [2] Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California 94143, USA [3] Department of Microbiology and Immunology, University of California San Francisco, San Francisco, California 94143, USA [4] The G.W. Hooper Foundation, University of California San Francisco, San Francisco, California 94143, USA.
Frances M
FM
Brodsky
D000199:Actins / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002465:Cell Movement; D033922:Clathrin Heavy Chains / Q000378:metabolism; D033941:Clathrin Light Chains / Q000378:metabolism*; D004705:Endocytosis; D005260:Female; D022001:Focal Adhesions / Q000378:metabolism; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D006367:HeLa Cells; D006801:Humans; D019012:Integrin beta1 / Q000378:metabolism; D008954:Models, Biological; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D014162:Transfection; D014327:Trophoblasts / Q000166:cytology* / Q000378:metabolism; D015854:Up-Regulation; D014945:Wound Healing
D000199:Actins; D033941:Clathrin Light Chains; D019012:Integrin beta1; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D033922:Clathrin Heavy Chains