Messenger RNA in HeLa cells

Kinetics of formation and decay

Robert H. Singer, Sheldon Penman

Research output: Contribution to journalArticle

256 Citations (Scopus)

Abstract

The polyadenylic acid-containing messenger RNA fraction of HeLa cells was measured by its affinity for oligedeoxythymidylate cellulose. Both the kinetics of initial labeling and the decay after a brief pulse of incorporation were examined. The kinetics of decay are complex, but can be approximated by assuming two populations; a short-lived species with a half-life of seven hours and a long-lived component with a half-life of 24 hours. It is estimated that the short-lived material comprises 33% of total cellular mRNA, while the relatively stable species amounts to 67% of the steady-state mRNA content. The two mRNA components with different decay times were observed simultaneously in the same cell population by measuring decay of 24-hour old mRNA labeled with 14C and RNA briefly labeled with 3H. The old mRNA had only a 24-hour decay component, while the new mRNA was biphasic. The decay of old and new mRNA was also observed after RNA synthesis was inhibited with actinomycin. Again, old mRNA decayed more slowly than recently labeled material. However, both decay times are significantly shorter in the presence of actinomycin and correspond to half-lives of approximately 4 and 12 hours. There is a small but significant difference in sedimentation distribution of new and old mRNA, the old mRNA sedimenting more slowly than new material, suggesting that the more stable species has a lower average molecular weight. The steady-state content of mRNA in HeLa cells amounts to 5.5% of the ribosomal RNA, or more than twice the amount of messenger RNA estimated to be on hemoglobin-synthesizing polyribosomes.

Original languageEnglish (US)
Pages (from-to)321-334
Number of pages14
JournalJournal of Molecular Biology
Volume78
Issue number2
DOIs
StatePublished - Aug 5 1973
Externally publishedYes

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HeLa Cells
Messenger RNA
Dactinomycin
Half-Life
RNA
Poly A
Ribosomal RNA
Polyribosomes
Cellulose
Population
Hemoglobins
Molecular Weight

ASJC Scopus subject areas

  • Virology

Cite this

Messenger RNA in HeLa cells : Kinetics of formation and decay. / Singer, Robert H.; Penman, Sheldon.

In: Journal of Molecular Biology, Vol. 78, No. 2, 05.08.1973, p. 321-334.

Research output: Contribution to journalArticle

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abstract = "The polyadenylic acid-containing messenger RNA fraction of HeLa cells was measured by its affinity for oligedeoxythymidylate cellulose. Both the kinetics of initial labeling and the decay after a brief pulse of incorporation were examined. The kinetics of decay are complex, but can be approximated by assuming two populations; a short-lived species with a half-life of seven hours and a long-lived component with a half-life of 24 hours. It is estimated that the short-lived material comprises 33{\%} of total cellular mRNA, while the relatively stable species amounts to 67{\%} of the steady-state mRNA content. The two mRNA components with different decay times were observed simultaneously in the same cell population by measuring decay of 24-hour old mRNA labeled with 14C and RNA briefly labeled with 3H. The old mRNA had only a 24-hour decay component, while the new mRNA was biphasic. The decay of old and new mRNA was also observed after RNA synthesis was inhibited with actinomycin. Again, old mRNA decayed more slowly than recently labeled material. However, both decay times are significantly shorter in the presence of actinomycin and correspond to half-lives of approximately 4 and 12 hours. There is a small but significant difference in sedimentation distribution of new and old mRNA, the old mRNA sedimenting more slowly than new material, suggesting that the more stable species has a lower average molecular weight. The steady-state content of mRNA in HeLa cells amounts to 5.5{\%} of the ribosomal RNA, or more than twice the amount of messenger RNA estimated to be on hemoglobin-synthesizing polyribosomes.",
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