Overexpression of protein kinase G using adenovirus inhibits cyclin E transcription and mesangial cell cycle

Am J Physiol Renal Physiol. 2001 May;280(5):F851-9. doi: 10.1152/ajprenal.2001.280.5.F851.

Abstract

The cGMP-cGMP-dependent protein kinase (protein kinase G) system plays an important role in the pathogenesis of mesangial proliferative glomerulonephritis. However, the molecular mechanisms of the inhibitory effects of the cGMP-protein kinase G system in the cell cycle progression of mesangial cells are not well known. To determine the inhibitory pathway of cGMP-protein kinase G in cultured mesangial cells, we investigated the effects of cGMP- and adenovirus-mediated overexpression of protein kinase G on the promoter activities of cyclin E, cyclin D1, and cyclin A. 8-Bromo-cGMP (8-BrcGMP) and overexpression of protein kinase G reduced [(3)H]thymidine uptake, reduced the numbers of cells in S and G(2)/M phases, and decreased the phosphorylation of retinoblastoma (Rb) protein. 8-BrcGMP (10(-3) M), protein kinase G adenovirus (Ad-cGKIbeta; 10(10) plaque-forming units/ml), atrial natriuretic peptide (ANP), and C-type natriuretic peptide (CNP) inhibited the promoter activity of cyclin E to 49, 57, 77, and 78%, respectively. On the other hand, the promoter activities of cyclin D1 and cyclin A were not changed significantly. In Western blot analysis, 8-BrcGMP, Ad-cGKIbeta, ANP, and CNP also inhibited cyclin E protein expression dose and time dependently. The p44/p42 mitogen-activated protein kinase (MAPK) kinase 1-p44/p42 MAPK had no effect on cyclin E promoter activities, and the cGMP-protein kinase G pathway did not change MAPK activity. In conclusion, our findings suggest that the reduction of the cyclin E promoter activity that downregulates G(1)/S transition plays a dominant role in the cGMP- and protein kinase G-induced inhibition of mesangial cell proliferation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / genetics*
  • Animals
  • Cell Count
  • Cell Cycle / physiology
  • Cyclic GMP / biosynthesis
  • Cyclic GMP-Dependent Protein Kinases / biosynthesis*
  • Cyclin E / biosynthesis*
  • Cyclin E / genetics
  • Flow Cytometry
  • Genes, Reporter
  • Genetic Vectors
  • Glomerular Mesangium / cytology*
  • Glomerular Mesangium / enzymology
  • Glomerular Mesangium / metabolism*
  • Luciferases
  • Male
  • Mitogen-Activated Protein Kinases / metabolism
  • Plasmids
  • RNA, Messenger / biosynthesis
  • Rats
  • Rats, Sprague-Dawley
  • Thymidine / metabolism
  • Transcription, Genetic / genetics

Substances

  • Cyclin E
  • RNA, Messenger
  • Luciferases
  • Cyclic GMP-Dependent Protein Kinases
  • Mitogen-Activated Protein Kinases
  • Cyclic GMP
  • Thymidine