HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis-2

<p><b>Copyright information:</b></p><p>Taken from "HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis"</p><p>http://www.biomedcentral.com/1471-2121/8/50</p><p>BMC Cell Biology 2007;8():50-50.</p><p>Published online 28 Nov 2007</p><p>PMCID:PMC2222240.</p><p></p>ne. Grey boxes indicate the hydrophobic region. TM boxes indicate the locations of putative transmembrane domain. (B) Expression of wild-type or deleted mutants in COS-7 cells. COS-7 cells were transfected with the indicated constructs in the expression vectors. The cells were incubated with MitoTracker and fixed. Fluorescent images of GFP (green), MitoTracker (red) were taken using a confocal microscope. Merged fluorescent images of GFP and MitoTracker are shown. Other conditions are described in Materials and methods

HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis-5

<p><b>Copyright information:</b></p><p>Taken from "HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis"</p><p>http://www.biomedcentral.com/1471-2121/8/50</p><p>BMC Cell Biology 2007;8():50-50.</p><p>Published online 28 Nov 2007</p><p>PMCID:PMC2222240.</p><p></p>100 nM of staurosporine and 300 nM of actinomycin D, in respectively. The cells were than stained with propidium iodide and analyzed by FACS. The % of apoptotic cells from each analysis were presented on a bar graph

HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis-6

<p><b>Copyright information:</b></p><p>Taken from "HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis"</p><p>http://www.biomedcentral.com/1471-2121/8/50</p><p>BMC Cell Biology 2007;8():50-50.</p><p>Published online 28 Nov 2007</p><p>PMCID:PMC2222240.</p><p></p>d conserved substitutions by gray. (B) Schematic alignment shows the LETM1 domain structure. Analysis using Pfam revealed sequence similarity with the LETM1 domains. Scale is lengths proportional to their molecular masses. The locations of putative LETM domains are indicated by blank boxes. Homo sapiens gi18204589 ref AAH21208; Homo sapiens (HCCR-1) gi13624098 ref AAK34885; Mus musculus gi33416955 ref AAH55685; Drosophila melanogaster gi21626751 ref AAM68317; Drosophila melanogaster gi1749774 ref CAA71125; Molgula oculata, gi308969 ref AAC37181; Caenorhabditis elegans, gi17561658 ref NP_506382; Saccharomyces cerevisiae, gi6324546 ref NP_014615; Saccharomyces cerevisiae, gi1762146 ref AAB70096; Ashbya gossypii gi44980464 ref AAS50397; Ashbya gossypii, gi44980448 ref AAS50381; Neurospora crassa gi32420419 ref XP_330653; Arabidopsis thaliana, gi42562974 ref NP_176732

HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis-0

<p><b>Copyright information:</b></p><p>Taken from "HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis"</p><p>http://www.biomedcentral.com/1471-2121/8/50</p><p>BMC Cell Biology 2007;8():50-50.</p><p>Published online 28 Nov 2007</p><p>PMCID:PMC2222240.</p><p></p>d conserved substitutions by gray. (B) Schematic alignment shows the LETM1 domain structure. Analysis using Pfam revealed sequence similarity with the LETM1 domains. Scale is lengths proportional to their molecular masses. The locations of putative LETM domains are indicated by blank boxes. Homo sapiens gi18204589 ref AAH21208; Homo sapiens (HCCR-1) gi13624098 ref AAK34885; Mus musculus gi33416955 ref AAH55685; Drosophila melanogaster gi21626751 ref AAM68317; Drosophila melanogaster gi1749774 ref CAA71125; Molgula oculata, gi308969 ref AAC37181; Caenorhabditis elegans, gi17561658 ref NP_506382; Saccharomyces cerevisiae, gi6324546 ref NP_014615; Saccharomyces cerevisiae, gi1762146 ref AAB70096; Ashbya gossypii gi44980464 ref AAS50397; Ashbya gossypii, gi44980448 ref AAS50381; Neurospora crassa gi32420419 ref XP_330653; Arabidopsis thaliana, gi42562974 ref NP_176732

HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis-4

<p><b>Copyright information:</b></p><p>Taken from "HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis"</p><p>http://www.biomedcentral.com/1471-2121/8/50</p><p>BMC Cell Biology 2007;8():50-50.</p><p>Published online 28 Nov 2007</p><p>PMCID:PMC2222240.</p><p></p>en incubated for 2 days. The cells were fractionated into the nuclei (lane 1), post-mitochondria (lane 2), and mitochondria (lane3) fractions. Each 10 μg of proteins was separated by SDS-PAGE and analyzed by immunoblotting using antibodies to the GFP epitope (top panel), the mitochondrial protein VDAC-1 (bottom panel)

HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis-3

<p><b>Copyright information:</b></p><p>Taken from "HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis"</p><p>http://www.biomedcentral.com/1471-2121/8/50</p><p>BMC Cell Biology 2007;8():50-50.</p><p>Published online 28 Nov 2007</p><p>PMCID:PMC2222240.</p><p></p> boxes indicate the hydrophobic region. TM boxes indicate the locations of putative transmembrane domain. (B) Expression of deleted mutants in COS-7 cells. COS-7 cells were transfected with the indicated constructs in the expression vectors. Other conditions were same as in Figure 3B

HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis-1

<p><b>Copyright information:</b></p><p>Taken from "HCCR-1, a novel oncogene, encodes a mitochondrial outer membrane protein and suppresses the UVC-induced apoptosis"</p><p>http://www.biomedcentral.com/1471-2121/8/50</p><p>BMC Cell Biology 2007;8():50-50.</p><p>Published online 28 Nov 2007</p><p>PMCID:PMC2222240.</p><p></p>ultured on the pre-coated coverslip for overnight. COS-7 and MCF-7 cells were transiently transfected with the pEGFP-N1 vector (GFP) or the vector carrying the human HCCR-1 gene tagged with GFP at its COOH-terminus (HCCR-1/GFP). The cells were incubated with MitoTracker (Red), fixed with 4% paraformaldyhyde. HEK293/HCCR-1-V5 cells were stained with MitoTracker (Red) and fixed with 100% methanol for 1 min at -20°C. Mouse anti-V5 antibody (1:300 dilution) and goat anti-mouse IgG conjugated with HRP (5 μg/ml) were applied to the cells for 1 h at room temperature (HCCR-1-V5). The cells were mounted and examined using a confocal microscope. (B) HCCR-1 is expressed at the mitochondrial membrane in the HEK293 cell line. Subcellular fraction of HEK293/HCCR-1-V5 cell line. HEK293/HCCR-1-V5 cells were fractionated into the nuclei (lane 1), post-mitochondria (lane 2), and mitochondria (lane3) fractions by differential centrifugation. Isolated mitochondria were treated with 0.1 M NaCOand fractionated by centrifugation at 100,000 g into pellets (lane 4) and supernatant (lane 5). Proteins were analyzed by immunoblotting using antibodies to the V5 epitope (top panel), the mitochondrial membrane protein VDAC-1 (middle panel) and the mitochondrial matrix protein SOD-2 (bottom panel). (C) HCCR-1 topology on the mitochondrial outer membrane. Intact mitochondria were incubated for 30 min at room temperature without (lane 1) and with 50 μg/ml proteinase K (lane 2) or with 50 μg/ml proteinase K plus 1% Triton X-100 (lane 3). Each sample was precipitated with TCA and separated by SDS-PAGE and analyzed by immunoblotting with antibodies to V5 (top panel), to the outer membrane protein TOM40 (second panel) and to the inner membrane protein TIM23 (third panel), and to the matrix protein SOD-2 (bottom panel). (D) Schematic diagram shows the HCCR-1 topology at the outer mitochondrial membrane