title: Tyrosyl phosphorylated PAK1 regulates breast cancer cell motility in response to prolactin through filamin A.
abstract: The p21-activated serine-threonine kinase (PAK1) is activated by small GTPase-dependent and -independent mechanisms and regulates cell motility. Both PAK1 and the hormone prolactin (PRL) have been implicated in breast cancer by numerous studies. We have previously shown that the PRL-activated tyrosine kinase JAK2 (Janus tyrosine kinase 2) phosphorylates PAK1 in vivo and identified tyrosines (Tyr) 153, 201, and 285 in the PAK1 molecule as sites of JAK2 tyrosyl phosphorylation. Here, we have used human breast cancer T47D cells stably overexpressing PAK1 wild type or PAK1 Y3F mutant in which Tyr(s) 153, 201, and 285 were mutated to phenylalanines to demonstrate that phosphorylation of these three tyrosines are required for maximal PRL-dependent ruffling. In addition, phosphorylation of these three tyrosines is required for increased migration of T47D cells in response to PRL as assessed by two independent motility assays. Finally, we show that PAK1 phosphorylates serine (Ser) 2152 of the actin-binding protein filamin A to a greater extent when PAK1 is tyrosyl phosphorylated by JAK2. Down-regulation of PAK1 or filamin A abolishes the effect of PRL on cell migration. Thus, our data presented here bring some insight into the mechanism of PRL-stimulated motility of breast cancer cells.
authors: Department of Biological Sciences, University of Toledo, Toledo, OH 43606-3390, USA.
authors: Alan
authors: A
authors: Hammer
authors: Leah
authors: L
authors: Rider
authors: Peter
authors: P
authors: Oladimeji
authors: Leslie
authors: L
authors: Cook
authors: Quanwen
authors: Q
authors: Li
authors: Raymond R
authors: RR
authors: Mattingly
authors: Maria
authors: M
authors: Diakonova
mesh_terms: D001943:Breast Neoplasms / Q000201:enzymology* / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D002999:Clone Cells; D003285:Contractile Proteins / Q000378:metabolism*; D005260:Female; D064448:Filamins; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D053614:Janus Kinase 2 / Q000378:metabolism; D008840:Microfilament Proteins / Q000378:metabolism*; D008954:Models, Biological; D050505:Mutant Proteins / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D010768:Phosphoserine / Q000378:metabolism; D019000:Phosphotyrosine / Q000378:metabolism*; D011388:Prolactin / Q000494:pharmacology*; D054462:p21-Activated Kinases / Q000378:metabolism*
chemical_list: D003285:Contractile Proteins; D064448:Filamins; D008840:Microfilament Proteins; D050505:Mutant Proteins; D049452:Green Fluorescent Proteins; D010768:Phosphoserine; D019000:Phosphotyrosine; D011388:Prolactin; D053614:Janus Kinase 2; D054462:p21-Activated Kinases