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Functional mRNA can be generated by RNA polymerase III

S Gunnery1, M B Mathews

  • 1Cold Spring Harbor Laboratory, New York 11724, USA.

Molecular and Cellular Biology
|July 1, 1995
PubMed
Summary

Human cells can translate RNA polymerase III (pol III) transcripts into functional messenger RNA (mRNA), challenging the belief that only RNA polymerase II (pol II) produces mRNA. This suggests pol III may create cellular mRNAs.

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Molecular and cellular biology·2001

Area of Science:

  • Molecular Biology
  • Gene Expression
  • RNA Biology

Background:

  • Eukaryotic messenger RNA (mRNA) synthesis is primarily attributed to RNA polymerase II (pol II).
  • RNA polymerase I (pol I) synthesizes long ribosomal RNAs (rRNAs), and RNA polymerase III (pol III) produces transfer RNAs (tRNAs), 5S rRNA, and other small RNAs.
  • The functional specialization of these polymerases has been considered obligatory.

Purpose of the Study:

  • To investigate whether RNA polymerase III (pol III) transcripts can function as messenger RNA (mRNA).
  • To determine if pol III transcripts can be translated into functional proteins in human cells.
  • To assess the translational potential of an artificial pol III transcript encoding the HIV-1 Tat protein.

Main Methods:

  • Constructed a chimeric RNA molecule (pVA-Tat) by placing the HIV-1 tat gene under a pol III promoter.
  • Transcribed pVA-Tat in vitro and in vivo in HeLa cells.
  • Assessed Tat protein production and transactivation activity using a reporter construct; analyzed RNA capping, polyadenylation, and polysome association.

Main Results:

  • The artificial pol III transcript (VA-Tat RNA) was accurately transcribed and produced functional Tat protein.
  • VA-Tat RNA, lacking 5' cap and 3' poly(A) tail, associated with polysomes and initiated translation via scanning.
  • Pol III-generated Tat protein transactivated a reporter gene, demonstrating its functional capacity, albeit less efficiently than pol II-generated mRNA.

Conclusions:

  • Human cells are capable of utilizing pol III transcripts as functional mRNAs.
  • 5' capping and 3' polyadenylation are not essential for translation initiation, though they may enhance it.
  • These findings suggest that RNA polymerase III, and potentially RNA polymerase I, could synthesize cellular mRNAs.

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