DOI: 10.1091/mbc.e06-09-0831. A Novel AAK1 Splice Variant Functions at Multiple Steps of the Endocytic Pathway

A Novel AAK1 Splice Variant Functions at Multiple Steps of the Endocytic Pathway

10.1091/mbc.e06-09-0831

Crossref journal-article

American Society for Cell Biology (ASCB)

Molecular Biology of the Cell (1076)

Abstract

Phosphorylation is a critical step in regulating receptor transport through the endocytic pathway. AAK1 is a serine/threonine kinase that is thought to coordinate the recruitment of AP-2 to receptors containing tyrosine-based internalization motifs by phosphorylating the μ2 subunit. Here we have identified a long form of AAK1 (AAK1L) that contains an extended C-terminus that encodes an additional clathrin-binding domain (CBD2) consisting of multiple low-affinity interaction motifs. Protein interaction studies demonstrate that AAK1L CBD2 directly binds clathrin. However, in vitro kinase assays reveal little difference between AAK1 isoforms in their basal or clathrin-stimulated kinase activity toward the AP-2 μ2 subunit. However, overexpression of AAK1L CBD2 impairs transferrin endocytosis, confirming an endocytic role for AAK1. Surprisingly, CBD2 overexpression or AAK1 depletion by RNA interference significantly impairs transferrin recycling from the early/sorting endosome. These observations suggest that AAK1 functions at multiple steps of the endosomal pathway by regulating transferrin internalization and its rapid recycling back to the plasma membrane from early/sorting endosome.

Bibliography

Henderson, D. M., & Conner, S. D. (2007). A Novel AAK1 Splice Variant Functions at Multiple Steps of the Endocytic Pathway. Molecular Biology of the Cell, 18(7), 2698â2706.

Authors 2

Davin M. Henderson (first)
Sean D. Conner (additional)

References 49 Referenced 49

10.1083/jcb.120.1.37 / J. Cell Biol by Carter L. L. (1993)
10.1073/pnas.86.23.9289 / Proc. Natl. Acad. Sci. USA by Chin D. J. (1989)
10.1038/17618 / Nature by Christoforidis S. (1999)
10.1016/S0092-8674(02)00735-3 / Cell by Collins B. M. (2002)
10.1083/jcb.200108123 / J. Cell Biol by Conner S. D. (2002)
10.1038/nature01451 / Nature by Conner S. D. (2003)
10.1046/j.1398-9219.2003.0142.x / Traffic by Conner S. D. (2003)
10.1083/jcb.200304069 / J. Cell Biol by Conner S. D. (2003)
10.1083/jcb.144.6.1203 / J. Cell Biol by Cope M. J. (1999)
10.1016/S0076-6879(95)57026-8 / Methods Enzymol by Damke H. (1995)
10.1083/jcb.200106089 / J. Cell Biol by Engqvist-Goldstein A. E. (2001)
10.1083/jcb.141.3.611 / J. Cell Biol by Futter C. E. (1998)
10.1083/jcb.200211080 / J. Cell Biol by Ghosh P. (2003)
10.1074/jbc.275.2.1365 / J. Biol. Chem by Greener T. (2000)
10.1091/mbc.9.8.2217
10.1083/jcb.200304079 / J. Cell Biol by Jackson A. P. (2003)
10.1091/mbc.4.12.1251
10.1074/jbc.M312717200 / J. Biol. Chem by Keyel P. A. (2004)
10.1093/dnares/6.3.197 / DNA Res by Kikuno R. (1999)
10.1242/jcs.02548 / J. Cell Sci by Lee D. W. (2005)
10.1074/jbc.M408430200 / J. Biol. Chem by Legendre-Guillemin V. (2005)
10.1016/S0021-9258(18)45334-3 / J. Biol. Chem by Manfredi J. J. (1987)
10.1038/nrm1315 / Nat. Rev. Mol. Cell Biol by Maxfield F. R. (2004)
10.1083/jcb.121.6.1257 / J. Cell Biol by Mayor S. (1993)
10.1523/JNEUROSCI.20-23-08667.2000 / J. Neurosci by Morgan J. R. (2000)
10.1091/mbc.e06-05-0452
10.1016/0092-8674(92)90033-9 / Cell by Nathke I. S. (1992)
10.1126/science.7569928 / Science by Ohno H. (1995)
10.1016/S0960-9822(01)00240-8 / Curr. Biol by Olusanya O. (2001)
10.1091/mbc.e04-04-0355
10.1038/nature03571 / Nature by Pelkmans L. (2005)
10.1073/pnas.95.11.6187 / Proc. Natl. Acad. Sci. USA by Ren M. (1998)
10.1083/jcb.200111068 / J. Cell Biol by Ricotta D. (2002)
10.1083/jcb.150.5.1137 / J. Cell Biol by Sever S. (2000)
10.1016/S0021-9258(19)67819-1 / J. Biol. Chem by Sipe D. M. (1991)
10.1038/sj.embor.embor776 / EMBO Rep by Smythe E. (2003)
10.1083/jcb.132.1.21 / J. Cell Biol by Stoorvogel W. (1996)
10.1091/mbc.e05-03-0204
10.1091/mbc.10.8.2687
10.1073/pnas.97.3.1096 / Proc. Natl. Acad. Sci. USA by ter Haar E. (2000)
10.1078/S0171-9335(04)70037-0 / Eur. J. Cell Biol by Umeda A. (2000)
10.1038/378632a0 / Nature by Ungewickell E. (1995)
10.1091/mbc.01-07-0380
10.1091/mbc.e02-06-0338
10.1093/emboj/18.16.4383 / EMBO J by Wendland B. (1999)
10.1126/science.1074222 / Science by Wettey F. R. (2002)
10.1083/jcb.135.3.635 / J. Cell Biol by Wilde A. (1996)
10.1111/j.1600-0854.2005.00346.x / Traffic by Zhang C. X. (2005)
10.1073/pnas.0403175101 / Proc. Natl. Acad. Sci. USA by Zhang L. (2004)

Dates

Type	When
Created	18 years, 3 months ago (May 9, 2007, 9:04 p.m.)
Deposited	4 years ago (Aug. 13, 2021, 3:53 a.m.)
Indexed	2 weeks, 2 days ago (Aug. 21, 2025, 1:59 p.m.)
Issued	18 years, 2 months ago (July 1, 2007)
Published	18 years, 2 months ago (July 1, 2007)
Published Print	18 years, 2 months ago (July 1, 2007)

Funders 0

None

BibTeX

@article{Henderson_2007, title={A Novel AAK1 Splice Variant Functions at Multiple Steps of the Endocytic Pathway}, volume={18}, ISSN={1939-4586}, url={http://dx.doi.org/10.1091/mbc.e06-09-0831}, DOI={10.1091/mbc.e06-09-0831}, number={7}, journal={Molecular Biology of the Cell}, publisher={American Society for Cell Biology (ASCB)}, author={Henderson, Davin M. and Conner, Sean D.}, editor={Drubin, David}, year={2007}, month=jul, pages={2698–2706} }

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      "year": "2000",
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      "year": "1991",
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      "DOI": "10.1083/jcb.132.1.21",
      "volume": "132",
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      "given": "David",
      "family": "Drubin",
      "sequence": "additional",
      "affiliation": []
    }
  ],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2007,
        7
      ]
    ]
  },
  "references-count": 49,
  "journal-issue": {
    "issue": "7",
    "published-print": {
      "date-parts": [
        [
          2007,
          7
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1091/mbc.e06-09-0831"
  ],
  "URL": "http://dx.doi.org/10.1091/mbc.E06-09-0831",
  "relation": {},
  "ISSN": [
    "1059-1524",
    "1939-4586"
  ],
  "subject": [],
  "container-title-short": "MBoC",
  "published": {
    "date-parts": [
      [
        2007,
        7
      ]
    ]
  }
}