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RNA polymerase II is an essential mRNA polyadenylation factor

Nature volume 395, pages 93–96 (1998)Cite this article

Abstract

Production of messenger RNA in eukaryotic cells is a complex, multistep process. mRNA polyadenylation, or 3′ processing, requires several protein factors, including cleavage/polyadenylation-specificity factor (CPSF), cleavage-stimulation factor, two cleavage factors and poly(A) polymerase (reviewed in refs 1, 2). These proteins seem to be unnecessary for other steps in mRNA synthesis such as transcription and splicing, and factors required for these processes were not considered to be essential for polyadenylation. Nonetheless, these reactions may be linked so that they are effectively coordinated in vivo3,4,5,6,7,8,9. For example, the CTD carboxy-terminal domain of the largest subunit of RNA polymerase II (RNAP II) is required for efficient splicing and polyadenylation in vivo8, and CPSF is brought to a promoter by the transcription factor TFIID and transferred to RNAP II at the time of transcription initiation9. These findings suggest that polyadenylation factors can be recruited to an RNA 3′-processing signal by RNAP II, where they dissociate from the polymerase and initiate polyadenylation. Here we present results that extend this model by showing that RNAP II is actually required, in the absence of transcription, for 3′ processing in vitro.

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Figure 1: Phosphoserine and RNA polymerase II activate pre-mRNA 3′ cleavage.
Figure 2: Both IIA and IIO isoforms of RNA polymerase II can activate 3′ cleavage.
Figure 3: The CTD is necessary and sufficient to reconstitute 3′ cleavage.
Figure 4: RNA polymerase II is required for 3′ cleavage in nuclear extract.

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Acknowledgements

We thank C. Li for the RNAP II used in Fig. 1 and for discussion; X. Sun and D.Reinberg for purified RNAP IIB; K. G. K. Murthy for recombinant PAP and antibodies; R. Tacke for purified SR proteins; W. S. Dynan for the GST–CTD plasmid; L. Tora, C. Kedinger and M. Vigneron for mAb 7G5; X. H. Shi, Y. Chen and Y. Sun for help in preparing nuclear extracts; and J. Dahlberg for suggesting that arginine phosphate might function by mimicking a phosphoarginine-containing protein. This work was supported by a grant from the NIH to J.M.; Y.H. was partly supported by the Japanese Ministry of Education, Science, and Culture.

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  1. Yutaka Hirose

    Present address: Department of Biophysics, Cancer Research Institute, Kanazawa University, 13-1, Takaramachi, Kanazawa, Ishikawa, 920, Japan

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  1. Department of Biological Sciences, Columbia University, New York, 10027, New York, USA

    James L. Manley

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Correspondence to James L. Manley.

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Hirose, Y., Manley, J. RNA polymerase II is an essential mRNA polyadenylation factor. Nature 395, 93–96 (1998). https://doi.org/10.1038/25786

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