Bunched and Madm Function Downstream of Tuberous Sclerosis Complex to Regulate the Growth of Intestinal Stem Cells in Drosophila

Stem Cell Rev Rep. 2015 Dec;11(6):813-25. doi: 10.1007/s12015-015-9617-5.

Abstract

The Drosophila adult midgut contains intestinal stem cells that support homeostasis and repair. We show here that the leucine zipper protein Bunched and the adaptor protein Madm are novel regulators of intestinal stem cells. MARCM mutant clonal analysis and cell type specific RNAi revealed that Bunched and Madm were required within intestinal stem cells for proliferation. Transgenic expression of a tagged Bunched showed a cytoplasmic localization in midgut precursors, and the addition of a nuclear localization signal to Bunched reduced its function to cooperate with Madm to increase intestinal stem cell proliferation. Furthermore, the elevated cell growth and 4EBP phosphorylation phenotypes induced by loss of Tuberous Sclerosis Complex or overexpression of Rheb were suppressed by the loss of Bunched or Madm. Therefore, while the mammalian homolog of Bunched, TSC-22, is able to regulate transcription and suppress cancer cell proliferation, our data suggest the model that Bunched and Madm functionally interact with the TOR pathway in the cytoplasm to regulate the growth and subsequent division of intestinal stem cells.

Keywords: Bunched; Drosophila; Intestine; Madm; Stem cells; TSC-22; Tuberous sclerosis complex.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cell Proliferation / genetics*
  • DNA-Binding Proteins / genetics*
  • DNA-Binding Proteins / metabolism
  • Drosophila
  • Drosophila Proteins / biosynthesis
  • Drosophila Proteins / genetics*
  • Drosophila Proteins / metabolism
  • Intestines / cytology
  • Monomeric GTP-Binding Proteins / biosynthesis
  • Neuropeptides / biosynthesis
  • RNA Interference
  • RNA, Small Interfering / genetics
  • Ras Homolog Enriched in Brain Protein
  • Signal Transduction
  • Stem Cells / cytology*
  • Stem Cells / metabolism
  • TOR Serine-Threonine Kinases / metabolism
  • Tuberous Sclerosis / genetics*
  • Tuberous Sclerosis / metabolism
  • Tumor Suppressor Proteins / genetics*
  • Tumor Suppressor Proteins / metabolism

Substances

  • DNA-Binding Proteins
  • Drosophila Proteins
  • Madm protein, Drosophila
  • Neuropeptides
  • RNA, Small Interfering
  • Ras Homolog Enriched in Brain Protein
  • Rheb protein, Drosophila
  • Tumor Suppressor Proteins
  • bun protein, Drosophila
  • TOR Serine-Threonine Kinases
  • Monomeric GTP-Binding Proteins