Intracellular proadrenomedullin-derived peptides decorate the microtubules and contribute to cytoskeleton function

Endocrinology. 2008 Jun;149(6):2888-98. doi: 10.1210/en.2007-1763. Epub 2008 Mar 6.

Abstract

Adrenomedullin (AM) and proadrenomedullin N-terminal 20 peptide (PAMP) are secretory hormones, but it is not unusual to find them in intracellular compartments. Using yeast-2 hybrid technology, we found interactions between AM and several microtubule-associated proteins (MAPs), and between PAMP and tubulin. Expression of fluorescent-tagged AM and PAMP as well as immunofluorescence for the native peptides showed a complete decoration of the microtubules and colocalization with other MAPs. PAMP, but not AM, bound to tubulin in vitro and destabilized tubulin polymerization. Down-regulation of the gene coding for both AM and PAMP through small interfering RNA technology resulted in morphological changes, microtubule stabilization, increase in posttranslational modifications of tubulin such as acetylation and detyrosination, reduction in cell motility, and partial arrest at the G2 phase of the cell cycle, when compared with cells transfected with the same vector carrying a scrambled sequence. These results show that PAMP is a novel MAP, whereas AM may be exerting more subtle effects in regulating cytoskeleton function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma
  • Adrenomedullin / chemistry
  • Adrenomedullin / genetics
  • Adrenomedullin / physiology*
  • Animals
  • CHO Cells
  • COS Cells
  • Cell Line, Tumor
  • Chlorocebus aethiops
  • Cricetinae
  • Cricetulus
  • Cytoskeleton / physiology*
  • Haplorhini
  • Humans
  • Lung Neoplasms
  • Microtubules / physiology*
  • Peptide Fragments / physiology*
  • Protein Precursors / chemistry
  • Protein Precursors / genetics
  • Protein Precursors / physiology*

Substances

  • Peptide Fragments
  • Protein Precursors
  • proadrenomedullin
  • Adrenomedullin