MTERF1 regulates the oxidative phosphorylation activity and cell proliferation in HeLa cells

Abstract

The mitochondrial transcription termination factor (MTERF) family is a group of highly conserved DNA-binding proteins composed of four key members, MTERF1-4. To date, several studies have investigated the binding sites of MTERF1 on mitochondrial genome and the regulation of mitochondrial gene transcription, but the more intricate connection between mitochondrial genes transcription regulation, mitochondrial oxidative phosphorylation (OXPHOS), and cell proliferation is still poorly understood. In this study, we constructed overexpression and knockdown vectors of MTERF1 that were transfected into HeLa cells to investigate the functions of MTERF1. Results showed that although MTERF1 is a positive regulatory factor of mitochondrial genes transcription, it had no significant effect on the replication of mitochondrial DNA. Over-expression of MTERF1 increased mitochondrial oxidative phosphorylation activity and promoted ATP synthesis, cyclin D1 expression, and cell proliferation, while its knockdown inhibited ATP synthesis, decreased cyclin D1 expression, and slowed the cell growth. These results suggested that MTERF1 may promote cell proliferation by regulating oxidative phosphorylation activity in HeLa cells. Ultimately, these findings create a foundation for further and more conclusive studies on the physiological functions of MTERF family by providing novel insights into the potential mechanisms underlying cell proliferation regulation.

PGC-1α regulates the cell cycle through ATP and ROS in CH1 cells

“…After 48 h of culture, aminoglycoside antibiotic G418 (Gibco-BRL, Carlsbad, USA) was added into the medium at a final concentration of 600 μg/ml to select CH1 cells with stable expression of PGC-1α. PGC-1α expression was confirmed by Western blotting (Chen et al, 2014).…”

“…After being washed 3 times with PBST (PBS solution contain 0.1% Tween 20), the membrane was incubated with a secondary antibody (at 1:2000 (v/v) dilution) at room temperature for 2 h, and protein was detected with an enhanced chemiluminescence kit (Thermo scientific, Boston, USA). PGC-1α, CyclinD1, and CyclinB1 antibodies were purchased from Cell Signaling Technology Co. (CST, Boston, USA), and an antitubulin antibody was obtained from Beyotime Co. (Jiangsu, China) (Chen et al, 2014). To quantify relative protein expression, Western blots were analyzed using ImageJ software.…”

Bcl-2 delays cell cycle through mitochondrial ATP and ROS

“…NIH3T3 (3T3) and C3H cells were then infected with the PB vector or Bcl-2 virus for 48 hr, the stable expressing cells were screened by antibiotic G418 (Gibco-BRL, Carlsbad, USA, 10131035), and confirmed by Western blotting. 30 Detection of ATP and ROS during serum re-stimulation Cells were synchronized at G 0 /G 1 stage by contact inhibition (CI) and serum starvation (SS). After re-stimulation by re-plating or serum addition, cells were harvested every 2 hr starting from 8 hr to 20 hr, and stained with PI and analyzed for cell cycle by BD Accuri C6 flow cytometer (BD Biosciences, San Jose, CA).…”

Modulating p-AMPK/mTOR Pathway of Mitochondrial Dysfunction Caused by MTERF1 Abnormal Expression in Colorectal Cancer Cells

“…MTERF1 plays an important role in the occurrence and development of tumors. Overexpression of MTERF1 increased mitochondrial gene transcription and promoted ATP synthesis and cell proliferation in HeLa cells [ 18 ]. The mRNA level of MTERF1 is significantly increased in non-small cell lung cancer (NSCLC), and it is closely related to the improvement of the overall survival (OS) rate in patients with lung adenocarcinoma [ 19 ].…”