Membrane-associated RING-CH proteins associate with Bap31 and Target CD81 and CD44 to lysosomes

Eric Bartee, Craig A. Eyster, Kasinath Viswanathan, Mandana Mansouri, Julie G. Donaldson, Klaus Frueh

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Membrane-associated RING-CH (MARCH) proteins represent a family of transmembrane ubiquitin ligases modulating intracellular trafficking and turnover of transmembrane protein targets. While homologous proteins encoded by gamma-2 herpesviruses and leporipoxviruses have been studied extensively, limited information is available regarding the physiological targets of cellular MARCH proteins. To identify host cell proteins targeted by the human MARCH-VIII ubiquitin ligase we used stable isotope labeling of amino-acids in cell culture (SILAC) to monitor MARCH-dependent changes in the membrane proteomes of human fibroblasts. Unexpectedly, we observed that MARCH-VIII reduced the surface expression of Bap31, a chaperone that predominantly resides in the endoplasmic reticulum (ER). We demonstrate that Bap31 associates with the transmembrane domains of several MARCH proteins and controls intracellular transport of MARCH proteins. In addition, we observed that MARCH-VIII reduced the surface expression of the hyaluronic acid-receptor CD44 and both MARCH-VIII and MARCH-IV sequestered the tetraspanin CD81 in endo-lysosomal vesicles. Moreover, gene knockdown of MARCH-IV increased surface levels of endogenous CD81 suggesting a constitutive involvement of this family of ubiquitin ligases in the turnover of tetraspanins. Our data thus suggest a role of MARCH-VIII and MARCH-IV in the regulated turnover of CD81 and CD44, two ubiquitously expressed, multifunctional proteins.

Original languageEnglish (US)
Article numbere15132
JournalPLoS One
Volume5
Issue number12
DOIs
StatePublished - 2010

Fingerprint

lysosomes
Lysosomes
Membranes
ubiquitin
ligases
Proteins
proteins
Leporipoxvirus
Ligases
Ubiquitin
hyaluronic acid
isotope labeling
transmembrane proteins
proteome
endoplasmic reticulum
stable isotopes
fibroblasts
Tetraspanins
cell culture
Isotope Labeling

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Membrane-associated RING-CH proteins associate with Bap31 and Target CD81 and CD44 to lysosomes. / Bartee, Eric; Eyster, Craig A.; Viswanathan, Kasinath; Mansouri, Mandana; Donaldson, Julie G.; Frueh, Klaus.

In: PLoS One, Vol. 5, No. 12, e15132, 2010.

Research output: Contribution to journalArticle

Bartee, Eric ; Eyster, Craig A. ; Viswanathan, Kasinath ; Mansouri, Mandana ; Donaldson, Julie G. ; Frueh, Klaus. / Membrane-associated RING-CH proteins associate with Bap31 and Target CD81 and CD44 to lysosomes. In: PLoS One. 2010 ; Vol. 5, No. 12.
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