High complexity in the expression of the B′ subunit of protein phosphatase 2A0: Evidence for the existence of at least seven novel isoforms

Csilla Csortos, Stanislaw Zolnierowicz, Eva Bakó, Stephen D. Durbin, Anna De Paoli-Roach

Research output: Contribution to journalArticle

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Abstract

Association of the catalytic subunit (C2) with a variety of regulatory subunits is believed to modulate the activity and specificity of protein phosphatase 2A (PP2A). In this study we report the cloning and expression of a new family of B-subunit, the B′, associated with the PP2A0 form. Polymerase chain reactions and cDNA library screening have identified at least seven cDNA isotypes, designated α, β1, β2, β3, β4, γ, and δ. The different β subtypes appear to be generated by alternative splicing. The deduced amino acid sequences of the α, β2, β3, β4 and γ isoforms predict molecular weights of 57,600, 56,500, 60,900, 52,500, and 68,000, respectively. The proteins are 60-80% identical and differ mostly at their termini. Two of the isoforms, B′β3 and B′γ, contain a bipartite nuclear localization signal in their COOH terminus. No homology was found with other B- or B-related subunits. Northern analyses indicate a tissue-specific expression of the isoforms. Expression of B'a protein in Escherichia coli generated a polypeptide of ∼53 kDa, similar to the size of the B′ subunit present in the purified PP2A0. The recombinant protein was recognized by antibody raised against native B′ and interacted with the dimeric PP2A (A·C2) to generate a trimeric phosphatase. The deduced amino acid sequences of the B′ isoforms show significant homology to mammalian, fungal, and plant nucleotide sequences of unknown function present in the data bases. Notably, a high degree of homology (55-66%) was found with a yeast gene, RTS1, encoding a multicopy suppressor of a rox3 mutant. Our data indicate that at least seven B′ subunit isoforms may participate in the generation of a large number of PP2A0 holoenzymes that may be spatially and/or functionally targeted to different cellular processes.

Original languageEnglish
Pages (from-to)2578-2588
Number of pages11
JournalJournal of Biological Chemistry
Volume271
Issue number5
StatePublished - Feb 2 1996

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Phosphoprotein Phosphatases
Protein Isoforms
Protein Phosphatase 2
Amino Acid Sequence
Amino Acids
Nuclear Localization Signals
Holoenzymes
Gene encoding
Cloning
Escherichia coli Proteins
Polymerase chain reaction
Alternative Splicing
Gene Library
Phosphoric Monoester Hydrolases
Recombinant Proteins
Yeast
Escherichia coli
Organism Cloning
Catalytic Domain
Screening

ASJC Scopus subject areas

  • Biochemistry

Cite this

High complexity in the expression of the B′ subunit of protein phosphatase 2A0 : Evidence for the existence of at least seven novel isoforms. / Csortos, Csilla; Zolnierowicz, Stanislaw; Bakó, Eva; Durbin, Stephen D.; De Paoli-Roach, Anna.

In: Journal of Biological Chemistry, Vol. 271, No. 5, 02.02.1996, p. 2578-2588.

Research output: Contribution to journalArticle

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abstract = "Association of the catalytic subunit (C2) with a variety of regulatory subunits is believed to modulate the activity and specificity of protein phosphatase 2A (PP2A). In this study we report the cloning and expression of a new family of B-subunit, the B′, associated with the PP2A0 form. Polymerase chain reactions and cDNA library screening have identified at least seven cDNA isotypes, designated α, β1, β2, β3, β4, γ, and δ. The different β subtypes appear to be generated by alternative splicing. The deduced amino acid sequences of the α, β2, β3, β4 and γ isoforms predict molecular weights of 57,600, 56,500, 60,900, 52,500, and 68,000, respectively. The proteins are 60-80{\%} identical and differ mostly at their termini. Two of the isoforms, B′β3 and B′γ, contain a bipartite nuclear localization signal in their COOH terminus. No homology was found with other B- or B-related subunits. Northern analyses indicate a tissue-specific expression of the isoforms. Expression of B'a protein in Escherichia coli generated a polypeptide of ∼53 kDa, similar to the size of the B′ subunit present in the purified PP2A0. The recombinant protein was recognized by antibody raised against native B′ and interacted with the dimeric PP2A (A·C2) to generate a trimeric phosphatase. The deduced amino acid sequences of the B′ isoforms show significant homology to mammalian, fungal, and plant nucleotide sequences of unknown function present in the data bases. Notably, a high degree of homology (55-66{\%}) was found with a yeast gene, RTS1, encoding a multicopy suppressor of a rox3 mutant. Our data indicate that at least seven B′ subunit isoforms may participate in the generation of a large number of PP2A0 holoenzymes that may be spatially and/or functionally targeted to different cellular processes.",
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