Molecular cloning and analysis of a yeast protein phosphatase with an unusual amino-terminal region

Francesc Posas, Antonio Casamayor, Nuria Morral, Joaquín Ariño

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

DNA fragments containing structural characteristics found in Ser/Thr protein phosphatases were amplified by polymerase chain reaction from yeast genome. Amplification was carried out by using degenerate oligonucleotides encoding conserved sequences found in type 1, 2A, and 2B phosphatases. A 215-base pair amplification fragment was used to screen a size-selected library, and a positive clone was isolated and sequenced. Nucleotide sequencing revealed a 2076-base pair open reading frame encoding a 692-amino acid protein. The carboxyl half of the protein is structurally related to type 1 phosphatases and virtually identical with the sequence reported as PPZ1, whereas the amino-terminal half of the protein is unrelated to sequences found in other protein phosphatases. This region is very rich in Ser residues and presents a high number of basic amino acids. Therefore, the gene product, on the basis of its unique structure, would represent a novel class of protein phosphatase. The gene, which is located at chromosome XIII, is transcribed as a mRNA of about 2.7 kilobases, and the amount of message has been found to increase 3- to 4-fold during the culture. The product of the gene PPZ1 was identified by immunoblot analysis of cell extracts as a 75-kDa protein, and the amount of immunoreactive protein was increased in cells carrying multiple copies of the gene. Disruption of the gene resulted in viable cells, with no detectable phenotypic change, indicating that the gene is not essential for growth.

Original languageEnglish (US)
Pages (from-to)11734-11740
Number of pages7
JournalJournal of Biological Chemistry
Volume267
Issue number17
StatePublished - Jun 15 1992
Externally publishedYes

Fingerprint

Fungal Proteins
Cloning
Phosphoprotein Phosphatases
Molecular Cloning
Genes
Proteins
Phosphoric Monoester Hydrolases
Base Pairing
Amplification
Basic Amino Acids
Conserved Sequence
Cell Extracts
Polymerase chain reaction
Oligonucleotides
Open Reading Frames
Libraries
Chromosomes
Cell culture
Yeast
Nucleotides

ASJC Scopus subject areas

  • Biochemistry

Cite this

Molecular cloning and analysis of a yeast protein phosphatase with an unusual amino-terminal region. / Posas, Francesc; Casamayor, Antonio; Morral, Nuria; Ariño, Joaquín.

In: Journal of Biological Chemistry, Vol. 267, No. 17, 15.06.1992, p. 11734-11740.

Research output: Contribution to journalArticle

Posas, Francesc ; Casamayor, Antonio ; Morral, Nuria ; Ariño, Joaquín. / Molecular cloning and analysis of a yeast protein phosphatase with an unusual amino-terminal region. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 17. pp. 11734-11740.
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