Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

The Rab8 GTPase regulates apical protein localization in intestinal cells

Nature volume 448, pages 366–369 (2007)Cite this article

Abstract

A number of proteins are known to be involved in apical/basolateral transport of proteins in polarized epithelial cells1,2,3,4,5,6,7. The small GTP-binding protein Rab8 was thought to regulate basolateral transport in polarized kidney epithelial cells through the AP1B-complex-mediated pathway8,9. However, the role of Rab8 (Rab8A) in cell polarity in vivo remains unknown. Here we show that Rab8 is responsible for the localization of apical proteins in intestinal epithelial cells. We found that apical peptidases and transporters localized to lysosomes in the small intestine of Rab8-deficient mice. Their mislocalization and degradation in lysosomes led to a marked reduction in the absorption rate of nutrients in the small intestine, and ultimately to death. Ultrastructurally, a shortening of apical microvilli, an increased number of enlarged lysosomes, and microvillus inclusions in the enterocytes were also observed. One microvillus inclusion disease patient who shows an identical phenotype to Rab8-deficient mice expresses a reduced amount of RAB8 (RAB8A; NM_005370). Our results demonstrate that Rab8 is necessary for the proper localization of apical proteins and the absorption and digestion of various nutrients in the small intestine.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Accumulation of apical but not basolateral proteins in cytoplasm of Rab8-deficient intestinal epithelial cells.
Figure 2: The reduction of apical markers in the small intestine of the knockout mice causes a reduction of substrate uptake from the apical side.
Figure 3: Microvillus atrophy and inclusions in Rab8-deficient mice.
Figure 4: RAB8 is greatly reduced in intestinal epithelial cells of the microvillus inclusion disease patient.

Similar content being viewed by others

References

  1. Nelson, W. J. Adaptation of core mechanisms to generate cell polarity. Nature 422, 766–774 (2003)

    Article  ADS  CAS  Google Scholar 

  2. Schuck, S. & Simons, K. Polarized sorting in epithelial cells: raft clustering and the biogenesis of the apical membrane. J. Cell Sci. 117, 5955–5964 (2004)

    Article  CAS  Google Scholar 

  3. Rodriguez-Boulan, E., Musch, A. & Le Bivic, A. Epithelial trafficking: new routes to familiar places. Curr. Opin. Cell Biol. 16, 436–442 (2004)

    Article  CAS  Google Scholar 

  4. Matter, K. & Mellman, I. Mechanisms of cell polarity: sorting and transport in epithelial cells. Curr. Opin. Cell Biol. 6, 545–554 (1994)

    Article  CAS  Google Scholar 

  5. Zerial, M. & McBride, H. Rab proteins as membrane organizers. Nature Rev. Mol. Cell Biol. 2, 107–117 (2001)

    Article  CAS  Google Scholar 

  6. Sabatini, D. D. In awe of subcellular complexity: 50 years of trespassing boundaries within the cell. Annu. Rev. Cell Dev. Biol. 21, 1–33 (2005)

    Article  CAS  Google Scholar 

  7. Louvard, D., Kedinger, M. & Hauri, H. P. The differentiating intestinal epithelial cell: establishment and maintenance of functions through interactions between cellular structures. Annu. Rev. Cell Biol. 8, 157–195 (1992)

    Article  CAS  Google Scholar 

  8. Huber, L. A. et al. Rab8, a small GTPase involved in vesicular traffic between the TGN and the basolateral plasma membrane. J. Cell Biol. 123, 35–45 (1993)

    Article  CAS  Google Scholar 

  9. Ang, A. L. et al. The Rab8 GTPase selectively regulates AP-1B-dependent basolateral transport in polarized Madin-Darby canine kidney cells. J. Cell Biol. 163, 339–350 (2003)

    Article  CAS  Google Scholar 

  10. Rodriguez, C. I. et al. High-efficiency deleter mice show that FLPe is an alternative to Cre-loxP. Nature Genet. 25, 139–140 (2000)

    Article  CAS  Google Scholar 

  11. Ishida, Y. & Leder, P. RET: a poly A-trap retrovirus vector for reversible disruption and expression monitoring of genes in living cells. Nucleic Acids Res. 27, e35 (1999)

    Article  CAS  Google Scholar 

  12. Homareda, H., Nagano, Y. & Matsui, H. Immunochemical identification of exposed regions of the Na+,K+-ATPase α-subunit. FEBS Lett. 327, 99–102 (1993)

    Article  CAS  Google Scholar 

  13. Umesaki, Y., Tohyama, K. & Mutai, M. Biosynthesis of microvillus membrane-associated glycoproteins of small intestinal epithelial cells in germ-free and conventionalized mice. J. Biochem. 92, 373–379 (1982)

    Article  CAS  Google Scholar 

  14. Saito, H. et al. Cloning and characterization of a rat H+/peptide cotransporter mediating absorption of beta-lactam antibiotics in the intestine and kidney. J. Pharmacol. Exp. Ther. 275, 1631–1637 (1995)

    CAS  PubMed  Google Scholar 

  15. Koike, M. et al. Participation of autophagy in storage of lysosomes in neurons from mouse models of neuronal ceroid-lipofuscinoses (Batten disease). Am. J. Pathol. 167, 1713–1728 (2005)

    Article  CAS  Google Scholar 

  16. Nakamura, N. et al. Association of mouse sorting nexin 1 with early endosomes. J. Biochem. 130, 765–771 (2001)

    Article  CAS  Google Scholar 

  17. Tamai, I. et al. The predominant contribution of oligopeptide transporter PepT1 to intestinal absorption of beta-lactam antibiotics in the rat small intestine. J. Pharm. Pharmacol. 49, 796–801 (1997)

    Article  CAS  Google Scholar 

  18. Madison, B. B. et al. cis elements of the villin gene control expression in restricted domains of the vertical (crypt) and horizontal (duodenum, cecum) axes of the intestine. J. Biol. Chem. 277, 33275–33283 (2002)

    Article  CAS  Google Scholar 

  19. Davidson, G. P., Cutz, E., Hamilton, J. R. & Gall, D. G. Familial enteropathy: a syndrome of protracted diarrhea from birth, failure to thrive, and hypoplastic villus atrophy. Gastroenterology 75, 783–790 (1978)

    Article  CAS  Google Scholar 

  20. Phillips, A. D., Jenkins, P., Raafat, F. & Walker-Smith, J. A. Congenital microvillous atrophy: specific diagnostic features. Arch. Dis. Child. 60, 135–140 (1985)

    Article  CAS  Google Scholar 

  21. Harada, A. et al. Altered microtubule organization in small-calibre axons of mice lacking tau protein. Nature 369, 488–491 (1994)

    Article  ADS  CAS  Google Scholar 

  22. Sakai, K. & Miyazaki, J. A transgenic mouse line that retains Cre recombinase activity in mature oocytes irrespective of the cre transgene transmission. Biochem. Biophys. Res. Commun. 237, 318–324 (1997)

    Article  CAS  Google Scholar 

  23. Schwenk, F., Baron, U. & Rajewsky, K. A cre-transgenic mouse strain for the ubiquitous deletion of loxP-flanked gene segments including deletion in germ cells. Nucleic Acids Res. 23, 5080–5081 (1995)

    Article  CAS  Google Scholar 

  24. Harada, A., Sobue, K. & Hirokawa, N. Developmental changes of synapsin I subcellular localization in rat cerebellar neurons. Cell Struct. Funct. 15, 329–342 (1990)

    Article  CAS  Google Scholar 

  25. Harada, A. et al. MAP2 is required for dendrite elongation, PKA anchoring in dendrites, and proper PKA signal transduction. J. Cell Biol. 158, 541–549 (2002)

    Article  CAS  Google Scholar 

  26. Sato, M. et al. Caenorhabditis elegans RME-6 is a novel regulator of RAB-5 at the clathrin-coated pit. Nature Cell Biol. 7, 559–569 (2005)

    Article  CAS  Google Scholar 

  27. Kagitani, K. et al. Hypophosphatemic rickets accompanying congenital microvillous atrophy. J. Bone Miner. Res. 13, 1946–1952 (1998)

    Article  CAS  Google Scholar 

  28. Sasaki, T. et al. Zoom endoscopic evaluation of rejection in living-related small bowel transplantation. Transplantation 73, 560–564 (2002)

    Article  Google Scholar 

Download references

Acknowledgements

We thank M. Takano, T. Horie, R. Hirai, Y. Okada, C. Ohsawa, M. Sugiura, H. Hata and R. Ishida for assistance with cell culture, uptake studies, animal care, and microscopy, H. Gomi, T. Izumi, M. Komachi, C. Mogi, H. Tomura, F. Okajima, N. Shihara and T. Suzuki for teaching us various techniques, Y. Wada, Y. Uchiyama, H. Homareda, K. Inui and Y. Umesaki for providing antibodies, J. Miyazaki for providing the transgenic mice, S. Mitani for providing rab-8 knockout worms, and Y. Ishida for providing the construct. We thank the affected individuals and their families for providing the blood samples. The Lamp2 monoclonal antibody was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by The University of Iowa. We also thank H.-P. Zimmer, A. Ballauff and T. Berger for providing DNA samples of the patients. This work was supported by grants-in-aid and the 21st century Center of Excellence Program from the Japanese Ministry of Education, Culture, Sports, Science and Technology to T.S., M.S., K.S. and A.H.

Author Contributions T.S. generated and analysed Rab8 knockout mice. S.M. took care of the microvillus inclusion disease patient and collected the blood and small intestine samples. Y. Kato supervised the measurement of the absorption rate of nutrients in the intestine of the mice. K.S. and M.S. generated the transgenic C. elegans and photographed them. N.T. generated the chimeric mice. K.M. took care of the microvillus inclusion disease patient. K.O. supervised the treatment of the patient. Y. Kubo. measured the absorption rate of nutrients in the intestine. A.T. provided the instruments and supervised the absorption experiments. R.H. stained the tissue, arranged figures and wrote part of the manuscript. A.H. planned and supervised the experiments, performed morphological analyses, arranged figures and wrote the manuscript

Author information

Authors and Affiliations

  1. Department of Molecular and Cellullar Biology, Laboratory of Molecular Traffic, Institute for Molecular and Cellular Regulation, Gunma University, Gunma 371-8512, Japan,

    Takashi Sato, Ken Sato, Miyuki Sato, Reiko Harada & Akihiro Harada

  2. Department of Pediatrics, Osaka University Graduate School of Medicine, Osaka 565-0871, Japan,

    Sotaro Mushiake & Keiichi Ozono

  3. Division of Pharmaceutical Sciences, Graduate School of Natural Science and Technology, Kanazawa University, Kanazawa 920-1192, Japan,

    Yukio Kato, Yoshiyuki Kubo & Akira Tsuji

  4. Division of Transgenic Technology, Center for Animal Resources and Development (CARD), IRDA, Kumamoto University, Kumamoto 860-0811, Japan,

    Naoki Takeda

  5. Department of Pediatrics, Itami Municipal Hospital, Hyogo 664-8540, Japan,

    Kazunori Miki

Authors
  1. Takashi Sato
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sotaro Mushiake
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yukio Kato
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ken Sato
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Miyuki Sato
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Naoki Takeda
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Keiichi Ozono
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Kazunori Miki
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Yoshiyuki Kubo
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Akira Tsuji
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Reiko Harada
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Akihiro Harada
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Akihiro Harada.

Ethics declarations

Competing interests

Reprints and permissions information is available at www.nature.com/reprints. The authors declare no competing financial interests.

Supplementary information

Supplementary Information 1

This file contains Supplementary Figures S1-S14 and Legends, Supplementary Methods and additional references. (PDF 4222 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sato, T., Mushiake, S., Kato, Y. et al. The Rab8 GTPase regulates apical protein localization in intestinal cells. Nature 448, 366–369 (2007). https://doi.org/10.1038/nature05929

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature05929

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing