title: Intracellular proadrenomedullin-derived peptides decorate the microtubules and contribute to cytoskeleton function.
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.
authors.affiliation: Laboratory of Integrative and Medical Biophysics, National Cancer Institute, Bethesda, Maryland 20892, USA.
authors.forename: Dan L
authors.initials: DL
authors.lastname: Sackett
authors.forename: Laurent
authors.initials: L
authors.lastname: Ozbun
authors.forename: Enrique
authors.initials: E
authors.lastname: Zudaire
authors.forename: Lisa
authors.initials: L
authors.lastname: Wessner
authors.forename: John M
authors.initials: JM
authors.lastname: Chirgwin
authors.forename: Frank
authors.initials: F
authors.lastname: Cuttitta
authors.forename: Alfredo
authors.initials: A
authors.lastname: Martínez
mesh_terms: D000230:Adenocarcinoma; D053607:Adrenomedullin / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D000818:Animals; D016466:CHO Cells; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D003599:Cytoskeleton / Q000502:physiology*; D000882:Haplorhini; D006801:Humans; D008175:Lung Neoplasms; D008870:Microtubules / Q000502:physiology*; D010446:Peptide Fragments / Q000502:physiology*; D011498:Protein Precursors / Q000737:chemistry / Q000235:genetics / Q000502:physiology*
chemical_list: D010446:Peptide Fragments; D011498:Protein Precursors; C088633:proadrenomedullin; D053607:Adrenomedullin