Go to The Journal of Clinical Investigation
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Transfers
  • Advertising/recruitment
  • Contact
  • Current Issue
  • Past Issues
  • By specialty
    • COVID-19
    • Cardiology
    • Immunology
    • Metabolism
    • Nephrology
    • Oncology
    • Pulmonology
    • All ...
  • Videos
  • Collections
    • Recently published
    • Technical Advances
    • Clinical Medicine
    • Reviews
    • Editorials
    • Perspectives
    • Top read articles
  • JCI This Month
    • Current issue
    • Past issues

  • Current issue
  • Past issues
  • Specialties
  • Recently published
  • In-Press Preview
  • Concise Communication
  • Editorials
  • Viewpoint
  • Top read articles
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Transfers
  • Advertising/recruitment
  • Contact
Stiff matrix instigates type I collagen biogenesis by mammalian cleavage factor I complex-mediated alternative polyadenylation
Zijing Zhou, … , Ping Chen, Yong Zhou
Zijing Zhou, … , Ping Chen, Yong Zhou
Published January 14, 2020
Citation Information: JCI Insight. 2020;5(3):e133972. https://doi.org/10.1172/jci.insight.133972.
View: Text | PDF
Research Article Pulmonology

Stiff matrix instigates type I collagen biogenesis by mammalian cleavage factor I complex-mediated alternative polyadenylation

  • Text
  • PDF
Abstract

Alternative polyadenylation (APA) is a widespread and important mechanism in regulation of gene expression. Dysregulation of the 3′ UTR cleavage and polyadenylation represents a common characteristic among many disease states, including lung fibrosis. In this study, we investigated the role of mammalian cleavage factor I–mediated (CFIm-mediated) APA in regulating extracellular matrix production in response to mechanical stimuli from stiffened matrix simulating the fibrotic lungs. We found that stiff matrix downregulated expression of CFIm68, CFIm59 and CFIm25 subunits and promoted APA in favor of the proximal poly(A) site usage in the 3′ UTRs of type I collagen (COL1A1) and fibronectin (FN1) in primary human lung fibroblasts. Knockdown and overexpression of each individual CFIm subunit demonstrated that CFIm68 and CFIm25 are indispensable attributes of stiff matrix–induced APA and overproduction of COL1A1, whereas CFIm did not appear to mediate stiffness-regulated FN1 APA. Furthermore, expression of the CFIm subunits was associated with matrix stiffness in vivo in a bleomycin-induced mouse model of pulmonary fibrosis. These data suggest that stiff matrix instigates type I collagen biogenesis by selectively targeting mRNA transcripts for 3′ UTR shortening. The current study uncovered a potential mechanism for regulation of the CFIm complex by mechanical cues under fibrotic conditions.

Authors

Zijing Zhou, Jing Qu, Li He, Yi Zhu, Shan-Zhong Yang, Feng Zhang, Ting Guo, Hong Peng, Ping Chen, Yong Zhou

×

Figure 2

Stiff matrix promotes expression of COL1A1 and FN1 at both the mRNA and protein level.

Options: View larger image (or click on image) Download as PowerPoint
Stiff matrix promotes expression of COL1A1 and FN1 at both the mRNA and ...
Human lung fibroblast populations (n = 3) were cultured on soft and stiff polyacrylamide gels for 48 hours. (A) Relative levels of COL1A1 and FN1 mRNA were determined by real-time RT-PCR. GAPDH was used as internal reference control. Bar graphs represent (mean ± SD) 5 separate experiments. (B) Protein levels of COL1A1 and FN1 were determined by immunoblot. GAPDH was used as loading control. Relative density of COL1A1 and FN1 bands was normalized to GAPDH. Bar graphs represent (mean ± SD) 3 separate experiments. (C) Human lung fibroblasts were treated with 10 μg/mL actinomycin D, followed by culturing on soft and stiff polyacrylamide gels for 24 hours. Relative levels of COL1A1 and FN1 mRNA were determined by real-time RT-PCR at the indicated time points. 18S rRNA was used as internal reference control. Graphs represent mRNA decay and half-lives of COL1A1 and FN1 under soft and stiff matrix conditions after actinomycin D treatment and represent (mean ± SD) 3 separate experiments. (D) Cell viability after actinomycin D treatment for 24 hours under both soft and stiff matrix conditions was evaluated by live/dead cell viability assays. Scale bar: 20 μm. A 2-tailed Student’s t test was used for comparison between groups.
Follow JCI Insight:
Copyright © 2021 American Society for Clinical Investigation
ISSN 2379-3708

Sign up for email alerts