title: The haematopoietic GTPase RhoH modulates IL3 signalling through regulation of STAT activity and IL3 receptor expression.
abstract: BACKGROUND
  RhoH is a constitutively active member of the family of Rho GTPases. Its expression is restricted to the haematopoietic lineage, where it serves as a positive regulator for T cell selection and mast cell function and as a negative regulator for growth-related functions in other lineages. Here, we examined the activation of signal transducer and activator of transcription (STAT) proteins in response to stimulation with interleukin 3 (IL3).
  
  
  RESULTS
  Using the murine IL3-dependent cell line BaF3 we investigated the influence of RhoH protein expression levels on IL3-mediated cellular responses. RhoH overexpressing cells showed lower sensitivity to IL3 and decreased STAT5 activation. SiRNA-mediated repression of RhoH gene expression led to an increase in proliferation and STAT5 activity which correlated with an increased number of IL3 receptor α chain molecules, also known as CD123, expressed at the cell surface. Interestingly, these findings could be reproduced using human THP-1 cells as a model system for acute myeloid leukaemia, where low RhoH levels are known to be an unfavourable prognostic marker. Overexpression of RhoH on the other hand caused an induction of STAT1 activity and western blot analysis revealed that activated STAT1 is phosphorylated on Tyr701. STAT1 is known to induce apoptosis or cell cycle arrest and we detected an upregulation of cyclin-dependent kinase inhibitors (CDKI) p21Cip1 and p27Kip1 in RhoH overexpressing BaF3 cells.
  
  
  CONCLUSIONS
  We propose that RhoH functions as a negative regulator for IL3-induced signals through modulation of the JAK-STAT pathway. High levels of RhoH allow the IL3-dependent activation of STAT1 causing decreased proliferation through upregulation of p21Cip1 and p27Kip1. Low RhoH levels on the other hand led to an upregulation of IL3-dependent cell growth, STAT5 activity and an increase of CD123 surface expression, linking RhoH to a CD123/STAT5 phenotype that has been described in AML patients.
authors.affiliation: Department für Infektiologie, Medizinische Mikrobiologie und Hygiene, Ruprecht-Karls-Universität, Im Neuenheimer Feld 324, Heidelberg, Germany.
authors.forename: Mehtap S
authors.initials: MS
authors.lastname: Gündogdu
authors.forename: He
authors.initials: H
authors.lastname: Liu
authors.forename: Daniela
authors.initials: D
authors.lastname: Metzdorf
authors.forename: Dagmar
authors.initials: D
authors.lastname: Hildebrand
authors.forename: Michael
authors.initials: M
authors.lastname: Aigner
authors.forename: Klaus
authors.initials: K
authors.lastname: Aktories
authors.forename: Klaus
authors.initials: K
authors.lastname: Heeg
authors.forename: Katharina F
authors.initials: KF
authors.lastname: Kubatzky
mesh_terms: D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000378:metabolism; D007377:Interleukin-3 / Q000378:metabolism*; D051379:Mice; D016185:Receptors, Interleukin-3 / Q000378:metabolism*; D050791:STAT Transcription Factors / Q000378:metabolism*; D015398:Signal Transduction; D014157:Transcription Factors / Q000502:physiology*; D015854:Up-Regulation; D020741:rho GTP-Binding Proteins / Q000502:physiology*
chemical_list: C493957:Cdkn1a protein, mouse; C493961:Cdkn1b protein, mouse; D050759:Cyclin-Dependent Kinase Inhibitor p21; D007377:Interleukin-3; D016185:Receptors, Interleukin-3; C496930:RhoH protein, mouse; D050791:STAT Transcription Factors; D014157:Transcription Factors; D050760:Cyclin-Dependent Kinase Inhibitor p27; D020741:rho GTP-Binding Proteins