AMPK regulates mitotic spindle orientation through phosphorylation of myosin regulatory light chain

Mol Cell Biol. 2012 Aug;32(16):3203-17. doi: 10.1128/MCB.00418-12. Epub 2012 Jun 11.

Abstract

The proper orientation of the mitotic spindle is essential for mitosis; however, how these events unfold at the molecular level is not well understood. AMP-activated protein kinase (AMPK) regulates energy homeostasis in eukaryotes, and AMPK-null Drosophila mutants have spindle defects. We show that threonine(172) phosphorylated AMPK localizes to the mitotic spindle poles and increases when cells enter mitosis. AMPK depletion causes a mitotic delay with misoriented spindles relative to the normal division plane and a reduced number and length of astral microtubules. AMPK-depleted cells contain mitotic actin bundles, which prevent astral microtubule-actin cortex attachments. Since myosin regulatory light chain (MRLC) is an AMPK downstream target and mediates actin function, we investigated whether AMPK signals through MRLC to control spindle orientation. Mitotic levels of serine(19) phosphorylated MRLC (pMRLC(ser19)) and spindle pole-associated pMRLC(ser19) are abolished when AMPK function is compromised, indicating that AMPK is essential for pMRLC(ser19) spindle pole activity. Phosphorylation of AMPK and MRLC in the mitotic spindle is dependent upon calcium/calmodulin-dependent protein kinase kinase (CamKK) activity in LKB1-deficient cells, suggesting that CamKK regulates this pathway when LKB1 function is compromised. Taken together, these data indicate that AMPK mediates spindle pole-associated pMRLC(ser19) to control spindle orientation via regulation of actin cortex-astral microtubule attachments.

Publication types

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

MeSH terms

  • AMP-Activated Protein Kinases / metabolism*
  • Actins / chemistry
  • Animals
  • Cell Line, Tumor
  • Depsipeptides / pharmacology
  • HeLa Cells
  • Homeostasis
  • Humans
  • Image Processing, Computer-Assisted
  • Microscopy, Confocal / methods
  • Microscopy, Fluorescence / methods
  • Mitosis
  • Mutation
  • Myosin Light Chains / metabolism*
  • Phosphorylation
  • Spindle Apparatus / metabolism*

Substances

  • Actins
  • Depsipeptides
  • Myosin Light Chains
  • jasplakinolide
  • AMP-Activated Protein Kinases