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Characterization of cleavage and polyadenylation specificity factor and cloning of its 100-kilodalton subunit

Jenny, A. and Hauri, H. P. and Keller, W.. (1994) Characterization of cleavage and polyadenylation specificity factor and cloning of its 100-kilodalton subunit. Molecular and Cellular Biology, 14 (12). pp. 8183-8190.

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Official URL: http://edoc.unibas.ch/dok/A5257790

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Abstract

During the formation of the 3' ends of mRNA, the cleavage and polyadenylation specificity factor (CPSF) is required for 3' cleavage of the transcript as well as for subsequent polyadenylation. Using peptide sequences from a tryptic digest, we have cloned the 100-kDa subunit of CPSF. This subunit is a novel protein showing no homology to any known polypeptide in databases. Polyclonal antibodies against the C terminus of the protein inhibit the polyadenylation reaction. Polyclonal and monoclonal antibodies were used to characterize the composition of CPSF. Immunoprecipitations of CPSF from HeLa cell extracts and from labeled chromatographic fractions show the coprecipitation of all four subunits of 160, 100, 73, and 30 kDa. Proteins of 160 and 30 kDa that are specifically cross-linked to precursor RNA by UV irradiation were identified as CPSF subunits by immunoprecipitation. Immunofluorescent detection of CPSF in HeLa cells localized it in the nucleoplasm, excluding cytoplasm and nucleolar structures.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Former Organization Units Biozentrum > Cell Biology (Keller)
05 Faculty of Science > Departement Biozentrum > Former Organization Units Biozentrum > Pharmacology/Neurobiology (Hauri)
UniBasel Contributors:Hauri, Hans-Peter and Keller, Walter
Item Type:Article, refereed
Article Subtype:Research Article
Bibsysno:Link to catalogue
Publisher:American Society for Microbiology
ISSN:0270-7306
e-ISSN:1098-5549
Note:Publication type according to Uni Basel Research Database: Journal article
Identification Number:
Last Modified:07 Nov 2017 15:19
Deposited On:22 Mar 2012 13:18

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