Numb directs the subcellular localization of EAAT3 through binding the YxNxxF motif.

Abstract:

:Excitatory amino acid transporter type 3 (EAAT3, also known as SLC1A1) is a high-affinity, Na(+)-dependent glutamate carrier that localizes primarily within the cell and at the apical plasma membrane. Although previous studies have reported proteins and sequence regions involved in EAAT3 trafficking, the detailed molecular mechanism by which EAAT3 is distributed to the correct location still remains elusive. Here, we identify that the YVNGGF sequence in the C-terminus of EAAT3 is responsible for its intracellular localization and apical sorting in rat hepatoma cells CRL1601 and Madin-Darby canine kidney (MDCK) cells, respectively. We further demonstrate that Numb, a clathrin adaptor protein, directly binds the YVNGGF motif and regulates the localization of EAAT3. Mutation of Y503, N505 and F508 within the YVNGGF motif to alanine residues or silencing Numb by use of small interfering RNA (siRNA) results in the aberrant localization of EAAT3. Moreover, both Numb and the YVNGGF motif mediate EAAT3 endocytosis in CRL1601 cells. In summary, our study suggests that Numb is a pivotal adaptor protein that mediates the subcellular localization of EAAT3 through binding the YxNxxF (where x stands for any amino acid) motif.

journal_name

J Cell Sci

journal_title

Journal of cell science

authors

Su JF,Wei J,Li PS,Miao HH,Ma YC,Qu YX,Xu J,Qin J,Li BL,Song BL,Xu ZP,Luo J

doi

10.1242/jcs.185496

subject

Has Abstract

pub_date

2016-08-15 00:00:00

pages

3104-14

issue

16

eissn

0021-9533

issn

1477-9137

pii

jcs.185496

journal_volume

129

pub_type

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