Journal: Frontiers in Cell and Developmental Biology

Article Title: Plasmalogens and Photooxidative Stress Signaling in Myxobacteria, and How it Unmasked CarF/TMEM189 as the Δ1′-Desaturase PEDS1 for Human Plasmalogen Biosynthesis

doi: 10.3389/fcell.2022.884689

Figure Lengend Snippet: The AlphaFold2 predicted structure of human TMEM189 compared to the crystal structure of SCD. Left: Crystal structure determined for mouse stearoyl-CoA desaturase/SCD (PDB accession code; 6WF2). Right: Single-chain AlphaFold2 structure of human TMEM189 ( https://alphafold.ebi.ac.uk/entry/A5PLL7 ). The grey box highlights the transmembrane region with four helices in each structure. Shown below are close-ups of: (left) the diiron centre (dark spheres) and the nine coordinating His and one Asn (through a water molecule) in the SCD structure (a portion of the substrate in the active site is shown by thin lines); (right) the conserved His (magenta), Asp and Phe (both yellow) essential for function, and two conserved histidines (green) not required for activity in human TMEM189 (see text).

Article Snippet: Indeed, Kua homologues figured only as a UEV1 localization domain in a protein family PF10520 described as Kua-UEV1_localn; and their true function remained elusive even after a Tmem189 knockout mouse was generated in a large-scale international mouse phenotyping consortium and several associated phenotypes, such as decreased body mass, and eye, bone and blood abnormalities were described ( ).

Techniques: Activity Assay

Journal: Biochemistry and Biophysics Reports

Article Title: Blockage of TMEM189 induces G2/M arrest and inhibits the growth of breast tumors

doi: 10.1016/j.bbrep.2024.101744

Figure Lengend Snippet: TMEM189 expression level is associated with cancer malignancy (A) TMEM189 mRNA levels of TNBC and non-TNBC patients's tumor tissues from the Metabric datasets. (B) TMEM189 mRNA levels in three tumor stages of TNBC samples from the Metabric dataset. (C) The overall survival probability analysis for 320 TNBC patients from the Metabric dataset by TMEM189 mRNA level. (D–F) Overall survival analysis of The Human Protein Atlas cohorts of patients with breast, renal, and liver cancer. (G) Western blotting analysis for TMEM189 expression in various breast, liver, and pancreatic cancer cell lines. * p < 0.05, ** p < 0.01 and *** p < 0.001.

Article Snippet: Antibodies were as follows: TMEM189 (A8380, Abclonal), Tubulin (AF0001, Beyotime), Cyclin B1 (R23324, Zenbio), CDK1 (R23884, Zenbio), pCDK1 (310063, Zenbio).

Techniques: Expressing, Western Blot

Journal: Biochemistry and Biophysics Reports

Article Title: Blockage of TMEM189 induces G2/M arrest and inhibits the growth of breast tumors

doi: 10.1016/j.bbrep.2024.101744

Figure Lengend Snippet: TMEM189 promotes cancer cell growth and migration (A) HepG2, MCF7, and Capan1 cells stably expressing either scramble or human TMEM189 shRNA, and 4T1 cells stably expressing either scramble or mouse TMEM189 shRNA were subjected to quantitative real-time PCR to detect knockdown efficiency (n = 3 per group). (B) HepG2 cells stably expressing either scramble or human TMEM189 shRNA, and 4T1 cells stably expressing either scramble or mouse TMEM189 shRNA were subjected to Western blotting. (C) The cells as indicated were subjected to cell viability assay as described in Method (n = 3 per group). (D) The cells as indicated were subjected to cell colony formation assay as described in Method. The whole well of 6- well plate was presented. (E) Representative images of Transwell assay (Magnification, 10×. Scale bar, 100 μm). (F) Quantitation of (E) (n = 5 per group). (G) HepG2 cells stably expressing either empty vector (EV) or human TMEM189 were subjected to Western blotting. (H) Cell viability assay was performed for HepG2 cells stably expressing either empty vector (EV) or human TMEM189. (I) Colony formation assay was performed for HepG2 cells stably expressing either empty vector (EV) or human TMEM189. The whole well of 6- well plate was presented. (J–K) The migration ability of HepG2 cells stably expressing either empty vector (EV) or human TMEM189 was assessed by wound-healing assay. Photographs were taken at 0 and 48 h following the initial scratch. Migration rates were quantified by measuring three different wound areas (n = 3) (Magnification, 10×. Scale bar, 100 μm). Three separate experiments were performed. (*** p < 0.001). (L) Transwell assay was performed for HepG2 cells stably expressing either empty vector (EV) or human TMEM189. (Magnification, 10×. Scale bar, 100 μm).Data were shown as the means ± SEM. * p < 0.05, ** p < 0.01 and *** p < 0.001.

Article Snippet: Antibodies were as follows: TMEM189 (A8380, Abclonal), Tubulin (AF0001, Beyotime), Cyclin B1 (R23324, Zenbio), CDK1 (R23884, Zenbio), pCDK1 (310063, Zenbio).

Techniques: Migration, Stable Transfection, Expressing, shRNA, Real-time Polymerase Chain Reaction, Knockdown, Western Blot, Viability Assay, Colony Assay, Transwell Assay, Quantitation Assay, Plasmid Preparation, Wound Healing Assay

Journal: Biochemistry and Biophysics Reports

Article Title: Blockage of TMEM189 induces G2/M arrest and inhibits the growth of breast tumors

doi: 10.1016/j.bbrep.2024.101744

Figure Lengend Snippet: TMEM189 deficiency inhibits cancer cell proliferation by inducing G2/M arrest (A-C) HepG2 cells stably expressing either scramble or TMEM189 shRNA were subjected to transcriptomic analysis. (A) Heatmap and (B) Volcano map presented a total of 1052 genes differentially expressed, including 345 upregulated and 707 downregulated genes. (C) KEGG pathway analysis of differentially expressed genes. Advanced bubble chart shows enrichment of differentially expressed genes in signaling pathways. (D–E) Flow cytometry analysis and quantification were performed to check cell cycle phases of HepG2 cells stably expressing either scramble or TMEM189 shRNA (n = 3). (F) The mRNA levels of G2/M arrest-related markers were detected by quantitative real-time PCR in HepG2 cells stably expressing either scramble or TMEM189 shRNA. (G) The protein levels of G2/M arrest-related markers were detected by Western blot in HepG2 cells stably expressing either scramble or TMEM189 shRNA. (H–I) Flow cytometry analysis and quantification were performed to check cell cycle phases of HepG2 cells stably expressing either empty vector (EV) or TMEM189 (n = 3). Data were shown as the means ± SEM. * p < 0.05, ** p < 0.01 and *** p < 0.001.

Article Snippet: Antibodies were as follows: TMEM189 (A8380, Abclonal), Tubulin (AF0001, Beyotime), Cyclin B1 (R23324, Zenbio), CDK1 (R23884, Zenbio), pCDK1 (310063, Zenbio).

Techniques: Stable Transfection, Expressing, shRNA, Protein-Protein interactions, Flow Cytometry, Real-time Polymerase Chain Reaction, Western Blot, Plasmid Preparation

Journal: Biochemistry and Biophysics Reports

Article Title: Blockage of TMEM189 induces G2/M arrest and inhibits the growth of breast tumors

doi: 10.1016/j.bbrep.2024.101744

Figure Lengend Snippet: Deletion of TMEM189 inhibits the growth of breast cancer in vivo (A-F) 4T1 cells stably expressing either scramble or mouse TMEM189 shRNA were subcutaneously injected on both sides of the dorsum of the BALB/c mice in a total volume of 100 μL (2 × 10 4 cells/mouse). (A) The growth rate of xenografted mice. (B) Tumor volumes (tumor volume = width 2 × length × 1/2). (C) Tumors were isolated and weighed at the endpoint of experiments. (D) Representative images of tumor sections stained with Ki67 antibody. (E) The percentage of Ki67 positive cells. (F) The mRNA levels of G2/M arrest markers were detected by RT-PCR for tumor samples (n = 8). (G) Schematic model of the role of TMEM189 in tumor proliferation. Data were shown as the means ± SEM. * p < 0.05, ** p < 0.01 and *** p < 0.001.

Article Snippet: Antibodies were as follows: TMEM189 (A8380, Abclonal), Tubulin (AF0001, Beyotime), Cyclin B1 (R23324, Zenbio), CDK1 (R23884, Zenbio), pCDK1 (310063, Zenbio).

Techniques: In Vivo, Stable Transfection, Expressing, shRNA, Injection, Isolation, Staining, Reverse Transcription Polymerase Chain Reaction