Expression of rat protein phosphatase 2C (IA) in Escherichia coli

Arabidopsis thaliana BAC T07M07 encoding the abscisic acid-insensitive 4 (ABI4) locus has been sequenced completely. It contains a 95,713-bp insert and 24 predicted genes. Most putative genes were confirmed by gel-based RNA profiling and a cluster of ABA-regulated genes was identified. One of the 24 genes, designatedPP2C5, encodes a putative protein phosphatase 2C. The encoded protein was expressed in Escherichia coli, and its enzyme activity in vitro was confirmed.[The sequence data described in this paper have been submitted to GenBank under accession no. AF085279.]

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TPD1 of Saccharomyces cerevisiae encodes a protein phosphatase 2C-like activity implicated in tRNA splicing and cell separation.

Molecular and Cellular Biology ◽

10.1128/mcb.14.6.3634 ◽

1994 ◽

Vol 14 (6) ◽

pp. 3634-3645 ◽

Cited By ~ 40

Author(s):

M K Robinson ◽

W H van Zyl ◽

E M Phizicky ◽

J R Broach

Keyword(s):

Escherichia Coli ◽

Saccharomyces Cerevisiae ◽

Protein Phosphatase ◽

Cell Separation ◽

Temperature Sensitive ◽

Wild Type ◽

Protein Phosphatase 2C ◽

Trna Splicing ◽

Multiple Phenotypes ◽

Higher Eukaryotes

The Saccharomyces cerevisiae TPD1 gene has been implicated in tRNA splicing because a tpd1-1 mutant strain accumulates unspliced precursor tRNAs at high temperatures (W. H. van Zyl, N. Wills, and J. R. Broach, Genetics 123:55-68, 1989). The wild-type TPD1 gene was cloned by complementation of the tpd1-1 mutation and shown to encode a protein with substantial homology to protein phosphatase 2C (PP2C) of higher eukaryotes. Expression of Tpd1p in Escherichia coli results in PP2C-like activity. Strains deleted for the TPD1 gene exhibit multiple phenotypes: temperature-sensitive growth, accumulation of unspliced precursor tRNAs, sporulation defects, and failure of cell separation during mitotic growth. On the basis of the presence of these observable phenotypes and the fact that Tpd1p accounts for a small percentage of the observed PP2C activity, we argue that Tpd1p is a unique member of the PP2C family.

Download Full-text

TPD1 of Saccharomyces cerevisiae encodes a protein phosphatase 2C-like activity implicated in tRNA splicing and cell separation

Molecular and Cellular Biology ◽

10.1128/mcb.14.6.3634-3645.1994 ◽

1994 ◽

Vol 14 (6) ◽

pp. 3634-3645

Author(s):

M K Robinson ◽

W H van Zyl ◽

E M Phizicky ◽

J R Broach

Keyword(s):

Escherichia Coli ◽

Saccharomyces Cerevisiae ◽

Protein Phosphatase ◽

Cell Separation ◽

Temperature Sensitive ◽

Wild Type ◽

Protein Phosphatase 2C ◽

Trna Splicing ◽

Multiple Phenotypes ◽

Higher Eukaryotes

The Saccharomyces cerevisiae TPD1 gene has been implicated in tRNA splicing because a tpd1-1 mutant strain accumulates unspliced precursor tRNAs at high temperatures (W. H. van Zyl, N. Wills, and J. R. Broach, Genetics 123:55-68, 1989). The wild-type TPD1 gene was cloned by complementation of the tpd1-1 mutation and shown to encode a protein with substantial homology to protein phosphatase 2C (PP2C) of higher eukaryotes. Expression of Tpd1p in Escherichia coli results in PP2C-like activity. Strains deleted for the TPD1 gene exhibit multiple phenotypes: temperature-sensitive growth, accumulation of unspliced precursor tRNAs, sporulation defects, and failure of cell separation during mitotic growth. On the basis of the presence of these observable phenotypes and the fact that Tpd1p accounts for a small percentage of the observed PP2C activity, we argue that Tpd1p is a unique member of the PP2C family.

Download Full-text