title: RTKN2 Induces NF-KappaB Dependent Resistance to Intrinsic Apoptosis in HEK Cells and Regulates BCL-2 Genes in Human CD4(+) Lymphocytes.
abstract: The gene for Rhotekin 2 (RTKN2) was originally identified in a promyelocytic cell line resistant to oxysterol-induced apoptosis. It is differentially expressed in freshly isolated CD4(+) T-cells compared with other hematopoietic cells and is down-regulated following activation of the T-cell receptor. However, very little is known about the function of RTKN2 other than its homology to Rho-GTPase effector, rhotekin, and the possibility that they may have similar roles. Here we show that stable expression of RTKN2 in HEK cells enhanced survival in response to intrinsic apoptotic agents; 25-hydroxy cholesterol and camptothecin, but not the extrinsic agent, TNFα. Inhibitors of NF-KappaB, but not MAPK, reversed the resistance and mitochondrial pro-apoptotic genes, Bax and Bim, were down regulated. In these cells, there was no evidence of RTKN2 binding to the GTPases, RhoA or Rac2. Consistent with the role of RTKN2 in HEK over-expressing cells, suppression of RTKN2 in primary human CD4(+) T-cells reduced viability and increased sensitivity to 25-OHC. The expression of the pro-apoptotic genes, Bax and Bim were increased while BCL-2 was decreased. In both cell models RTKN2 played a role in the process of intrinsic apoptosis and this was dependent on either NF-KappaB signaling or expression of downstream BCL-2 genes. As RTKN2 is a highly expressed in CD4(+) T-cells it may play a role as a key signaling switch for regulation of genes involved in T-cell survival.
authors.affiliation: Barwon Biomedical Research, Geelong Hospital, Barwon Health, Ryrie St, Geelong, Victoria, 3227, Australia. ; Metabolic Research Unit, School of Medicine and Institute for Technology Research and Innovation, Deakin University, Waurn Ponds, Victoria, 3217, Australia.
authors.forename: Fiona M
authors.initials: FM
authors.lastname: Collier
authors.affiliation: Barwon Biomedical Research, Geelong Hospital, Barwon Health, Ryrie St, Geelong, Victoria, 3227, Australia.
authors.forename: Andrea
authors.initials: A
authors.lastname: Loving
authors.affiliation: Department of Malignant Haematology and Stem Cell Transplantation, The Alfred Hospital, Melbourne, Victoria, 3181, Australia.
authors.forename: Adele J
authors.initials: AJ
authors.lastname: Baker
authors.affiliation: School of Medicine, Deakin University, Waurn Ponds, Victoria, 3217, Australia.
authors.forename: Janet
authors.initials: J
authors.lastname: McLeod
authors.affiliation: Metabolic Research Unit, School of Medicine and Institute for Technology Research and Innovation, Deakin University, Waurn Ponds, Victoria, 3217, Australia.
authors.forename: Ken
authors.initials: K
authors.lastname: Walder
authors.affiliation: Barwon Biomedical Research, Geelong Hospital, Barwon Health, Ryrie St, Geelong, Victoria, 3227, Australia.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Kirkland
keywords: Bcl-2 genes; NF-KappaB; T-cells; resistance to apoptosis; signaling