Myosin Vb Traffics P-Glycoprotein to the Apical Membrane of Intestinal Epithelial Cells

Sarah A. Dooley, Elena Kolobova, Andreanna Burman, Izumi Kaji, Jessica R. Digrazia, Rachel Stubler, Anna Goldstein, Charulekha Packirisamy, Alexander W. Coutts, Milena Saqui-Salces, Nan Gao, Melinda A. Engevik, Mitchell D. Shub, James R. Goldenring, Amy C. Engevik

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Background & Aims: The xenobiotic efflux pump P-glycoprotein is highly expressed on the apical membrane of the gastrointestinal tract, where it regulates the levels of intracellular substrates. P-glycoprotein is altered in disease, but the mechanisms that regulate the levels of P-glycoprotein are still being explored. The molecular motor myosin Vb (Myo5b) traffics diverse cargo to the apical membrane of intestinal epithelial cells. We hypothesized that Myo5b was responsible for the delivery of P-glycoprotein to the apical membrane of enterocytes. Methods: We used multiple murine models that lack functional Myo5b or the myosin binding partner Rab11a to analyze P-glycoprotein localization. Pig and human tissue were analyzed to determine P-glycoprotein localization in the setting of MYO5B mutations. Intestinal organoids were used to examine P-glycoprotein trafficking and to assay P-glycoprotein function when MYO5 is inhibited. Results: In mice lacking Myo5b or the binding partner Rab11a, P-glycoprotein was improperly trafficked and had decreased presence in the brush border of enterocytes. Immunostaining of a pig model lacking functional Myo5b and human biopsies from a patient with an inactivating mutation in Myo5b also showed altered localization of intestinal P-glycoprotein. Human intestinal organoids expressing the motorless MYO5B tail domain had colocalization with P-glycoprotein, confirming that P-glycoprotein was trafficked by MYO5B in human enterocytes. Inhibition of MYO5 in human intestinal cell lines and organoids resulted in decreased P-glycoprotein capacity. Additionally, inhibition of MYO5 in human colon cancer cells diminished P-glycoprotein activity and increased cell death in response to a chemotherapeutic drug. Conclusions: Collectively, these data demonstrate that Myo5b is necessary for the apical delivery of P-glycoprotein.

Original languageEnglish (US)
Pages (from-to)84-98.e9
JournalGastroenterology
Volume168
Issue number1
DOIs
StatePublished - Jan 2025

Bibliographical note

Publisher Copyright:
© 2025 AGA Institute

Keywords

  • Cell Trafficking
  • MDR1
  • Myosin Vb
  • P-Glycoprotein

PubMed: MeSH publication types

  • Journal Article

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