Kinetically distinct sorting pathways through the Golgi exhibit different requirements for Arf1

Michael Whitt, Michelle E. Cox, Rita Kansal, John Cox

Research output: Contribution to journalArticle

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Abstract

To investigate the role of cytoplasmic sequences in directing transmembrane protein trafficking through the Golgi, we analyzed the sorting of VSV tsO45 G fusions with either the native G cytoplasmic domain (G) or an alternative cytoplasmic tail derived from the chicken AE1-4 anion exchanger (GAE). At restrictive temperature GAE and G accumulated in the ER, and upon shifting the cells to permissive temperature both proteins folded and underwent transport through the Golgi. However, GAE and G did not form hetero-oligomers upon the shift to permissive temperature and they progressed through the Golgi with distinct kinetics. In addition, the transport of G through the proximal Golgi was Arf1 and COPI-dependent, while GAE progression through the proximal Golgi was Arf1 and COPI-independent. Although Arf1 did not regulate the sorting of GAE in the cis-Golgi, Arf1 did regulate the exit of GAE from the TGN. The trafficking of GAE through the Golgi was similar to that of the native AE1-4 anion exchanger, in that the progression of both proteins through the proximal Golgi was Arf1-independent, while both required Arf1 to exit the TGN. We propose that the differential recognition of cytosolic signals in membrane-spanning proteins by the Arf1-dependent sorting machinery may influence the rate at which cargo progresses through the Golgi. VSV tsO45 G with either the native G cytoplasmic domain (G) or an alternative cytoplasmic tail derived from an AE1 anion exchanger (GAE) progress through the Golgi with distinct kinetics. The transport of G through the proximal Golgi is Arf1-dependent, while GAE sorting in the proximal Golgi is Arf1-independent. We propose that the differential recognition of cytosolic signals in membrane cargo by the Arf1-dependent sorting machinery is an important factor in determining the rate of cargo transport through the Golgi.

Original languageEnglish (US)
Pages (from-to)267-283
Number of pages17
JournalTraffic
Volume16
Issue number3
DOIs
StatePublished - Jan 1 2015

Fingerprint

Erythrocyte Anion Exchange Protein 1
Coat Protein Complex I
Sorting
Temperature
Chloride-Bicarbonate Antiporters
Tail
Machinery
Protein Transport
Proteins
Chickens
Membrane Proteins
Membranes
Kinetics
Oligomers
Fusion reactions

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Kinetically distinct sorting pathways through the Golgi exhibit different requirements for Arf1. / Whitt, Michael; Cox, Michelle E.; Kansal, Rita; Cox, John.

In: Traffic, Vol. 16, No. 3, 01.01.2015, p. 267-283.

Research output: Contribution to journalArticle

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