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Updated: Jun 28, 2026
10:56
Polysome Fractionation and Analysis of Mammalian Translatomes on a Genome-wide Scale
Published on: May 17, 2014
Global and gene-specific translational regulation in rat lung development
Gail Otulakowski1, Wenming Duan, Hugh O'Brodovich
1Program in Physiology and Experimental Medicine, Hospital for Sick Children Research Institute, 555 University Avenue, Toronto, ON M5G1X8, Canada. gail.otulakowski@sickkids.ca
American Journal of Respiratory Cell and Molecular Biology
|October 28, 2008
Summary
Perinatal lung development involves translational control mechanisms. Key protein synthesis regulators shift activity around birth, impacting gene-specific protein expression for lung function.
Area of Science:
- Pulmonary physiology
- Molecular biology
- Developmental biology
Background:
- The perinatal lung undergoes significant physiological changes to transition to gas exchange.
- Protein synthesis is metabolically demanding and sensitive to environmental stress.
- Translational control mechanisms may regulate protein synthesis during lung development.
Purpose of the Study:
- To investigate the role of translational control mechanisms in the perinatal rat lung.
- To identify changes in protein synthesis regulation during the transition from fetal to postnatal life.
- To determine gene-specific translational regulation in the developing lung.
Main Methods:
- Immunoblotting to assess phosphorylation of key protein synthesis regulators.
- Immunohistochemistry to localize phospho-S6 staining in lung tissue.
- m(7)GTP-sepharose pulldown assays to analyze translation initiation factor complex formation.
- Polysome profiling to evaluate global translation activity.
- Microarray analysis, RT-PCR, and Northern blotting to identify and confirm gene-specific translational changes.
Main Results:
- Phosphorylation of mTOR pathway proteins (4E-BP, p70S6K, S6) decreased prenatally and increased postnatally.
- eIF4E binding shifted from its inhibitor (4E-BP) to its activator (4G) after birth.
- Global polysome abundance decreased late in gestation and increased on postnatal day 1.
- Gene-specific translational changes were observed, including increased IL-18 prenatally and decreased protein phosphatase 1 regulatory subunit 2 postnatally.
Conclusions:
- Translational control mechanisms are active in the perinatal rat lung.
- Postnatal lung development involves coordinated changes in global and gene-specific protein synthesis.
- Specific genes like IL-18 and protein phosphatase 1 regulatory subunit 2 exhibit unique translational regulation patterns during the perinatal period.
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