Analysis of expression patterns of protein phosphatase-1 and phosphatase-2A in rat and bovine lenses

David W Li, H. Xiang, U. Fass, X. Y. Zhang

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

PURPOSE. The reversible phosphorylation and dephosphorylation at the serine and threonine residues on proteins play distinct roles in regulating multiple cellular activities. Whereas the protein serine-threonine kinases have been well studied in the lens system, very little is known about the expression and function of the serine-threonine phosphatases. The present article reports the expression patterns of protein phosphatase (PP)-1 and -2A in adult rat and bovine lenses. METHODS. Total RNAs and proteins were extracted from the epithelial and fiber cells of rat and bovine lenses. RT-PCR and Northern blot analysis were used to detect the mRNA expression levels in the epithelial cells and different fractions of fiber cells of these two types of lenses. Western blot was used to examine the protein expression levels in these different samples. An enzymatic assay was used to detect the activity distribution of PP-1 and -2A in these samples. RESULTS. The mRNAs for the PP-1 catalytic subunit (PP-1cs) and PP-2A catalytic subunit (PP-2Acs) were expressed in both epithelial and fiber cells of rat and bovine lenses. A detailed examination of the expression patterns of the two mRNAs in different fractions of fiber cells revealed that the cortical fiber cells (F1) contain the highest level of PP-1cs and -2Acs mRNAs (similar to those in the epithelial cells) among different fractions of fiber cells. The levels of the two mRNAs were sequentially decreased in the next layers of fiber cells (F2 and F3) and became barely detectable in the inner layers of fiber cells (F4 and N). In contrast to the mRNA expression patterns, the PP-1cs protein was mainly found in the epithelial cells. Among different layers of fiber cells, only cortical (F1) fiber cells contained detectable level of PP-1cs protein (bovine lenses contained a relatively higher level of PP-1cs than rat lenses in this region). In the remaining fiber cells, the PP-1cs protein was hardly detectable in rat lenses and slightly detectable in bovine lenses. The PP-2Acs protein was detectable only in the lens epithelial cells. Enzymatic assays revealed that the distribution patterns of PP-1 and -2A activities were similar to those of PP-1cs and -2Acs proteins. Furthermore, PP-1 activity was approximately four to five times higher than PP-2A activity in the lens epithelial cells. CONCLUSIONS. This study demonstrates that active PP-1 and -2A are mainly distributed in the lens epithelial cells, with PP-1 as a major phosphatase. The mRNAs and proteins for PP-1cs and -2Acs are differentially expressed in the epithelial and fiber cells of rat and bovine lenses.

Original languageEnglish (US)
Pages (from-to)2603-2609
Number of pages7
JournalInvestigative Ophthalmology and Visual Science
Volume42
Issue number11
StatePublished - Oct 22 2001

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Protein Phosphatase 1
Phosphoric Monoester Hydrolases
Lenses
Phosphoprotein Phosphatases
Protein Phosphatase 2
Epithelial Cells
Messenger RNA
Proteins
Enzyme Assays
Crystallins
Protein-Serine-Threonine Kinases
Threonine
Northern Blotting
Serine
Catalytic Domain

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Analysis of expression patterns of protein phosphatase-1 and phosphatase-2A in rat and bovine lenses. / Li, David W; Xiang, H.; Fass, U.; Zhang, X. Y.

In: Investigative Ophthalmology and Visual Science, Vol. 42, No. 11, 22.10.2001, p. 2603-2609.

Research output: Contribution to journalArticle

Li, David W ; Xiang, H. ; Fass, U. ; Zhang, X. Y. / Analysis of expression patterns of protein phosphatase-1 and phosphatase-2A in rat and bovine lenses. In: Investigative Ophthalmology and Visual Science. 2001 ; Vol. 42, No. 11. pp. 2603-2609.
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abstract = "PURPOSE. The reversible phosphorylation and dephosphorylation at the serine and threonine residues on proteins play distinct roles in regulating multiple cellular activities. Whereas the protein serine-threonine kinases have been well studied in the lens system, very little is known about the expression and function of the serine-threonine phosphatases. The present article reports the expression patterns of protein phosphatase (PP)-1 and -2A in adult rat and bovine lenses. METHODS. Total RNAs and proteins were extracted from the epithelial and fiber cells of rat and bovine lenses. RT-PCR and Northern blot analysis were used to detect the mRNA expression levels in the epithelial cells and different fractions of fiber cells of these two types of lenses. Western blot was used to examine the protein expression levels in these different samples. An enzymatic assay was used to detect the activity distribution of PP-1 and -2A in these samples. RESULTS. The mRNAs for the PP-1 catalytic subunit (PP-1cs) and PP-2A catalytic subunit (PP-2Acs) were expressed in both epithelial and fiber cells of rat and bovine lenses. A detailed examination of the expression patterns of the two mRNAs in different fractions of fiber cells revealed that the cortical fiber cells (F1) contain the highest level of PP-1cs and -2Acs mRNAs (similar to those in the epithelial cells) among different fractions of fiber cells. The levels of the two mRNAs were sequentially decreased in the next layers of fiber cells (F2 and F3) and became barely detectable in the inner layers of fiber cells (F4 and N). In contrast to the mRNA expression patterns, the PP-1cs protein was mainly found in the epithelial cells. Among different layers of fiber cells, only cortical (F1) fiber cells contained detectable level of PP-1cs protein (bovine lenses contained a relatively higher level of PP-1cs than rat lenses in this region). In the remaining fiber cells, the PP-1cs protein was hardly detectable in rat lenses and slightly detectable in bovine lenses. The PP-2Acs protein was detectable only in the lens epithelial cells. Enzymatic assays revealed that the distribution patterns of PP-1 and -2A activities were similar to those of PP-1cs and -2Acs proteins. Furthermore, PP-1 activity was approximately four to five times higher than PP-2A activity in the lens epithelial cells. CONCLUSIONS. This study demonstrates that active PP-1 and -2A are mainly distributed in the lens epithelial cells, with PP-1 as a major phosphatase. The mRNAs and proteins for PP-1cs and -2Acs are differentially expressed in the epithelial and fiber cells of rat and bovine lenses.",
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N2 - PURPOSE. The reversible phosphorylation and dephosphorylation at the serine and threonine residues on proteins play distinct roles in regulating multiple cellular activities. Whereas the protein serine-threonine kinases have been well studied in the lens system, very little is known about the expression and function of the serine-threonine phosphatases. The present article reports the expression patterns of protein phosphatase (PP)-1 and -2A in adult rat and bovine lenses. METHODS. Total RNAs and proteins were extracted from the epithelial and fiber cells of rat and bovine lenses. RT-PCR and Northern blot analysis were used to detect the mRNA expression levels in the epithelial cells and different fractions of fiber cells of these two types of lenses. Western blot was used to examine the protein expression levels in these different samples. An enzymatic assay was used to detect the activity distribution of PP-1 and -2A in these samples. RESULTS. The mRNAs for the PP-1 catalytic subunit (PP-1cs) and PP-2A catalytic subunit (PP-2Acs) were expressed in both epithelial and fiber cells of rat and bovine lenses. A detailed examination of the expression patterns of the two mRNAs in different fractions of fiber cells revealed that the cortical fiber cells (F1) contain the highest level of PP-1cs and -2Acs mRNAs (similar to those in the epithelial cells) among different fractions of fiber cells. The levels of the two mRNAs were sequentially decreased in the next layers of fiber cells (F2 and F3) and became barely detectable in the inner layers of fiber cells (F4 and N). In contrast to the mRNA expression patterns, the PP-1cs protein was mainly found in the epithelial cells. Among different layers of fiber cells, only cortical (F1) fiber cells contained detectable level of PP-1cs protein (bovine lenses contained a relatively higher level of PP-1cs than rat lenses in this region). In the remaining fiber cells, the PP-1cs protein was hardly detectable in rat lenses and slightly detectable in bovine lenses. The PP-2Acs protein was detectable only in the lens epithelial cells. Enzymatic assays revealed that the distribution patterns of PP-1 and -2A activities were similar to those of PP-1cs and -2Acs proteins. Furthermore, PP-1 activity was approximately four to five times higher than PP-2A activity in the lens epithelial cells. CONCLUSIONS. This study demonstrates that active PP-1 and -2A are mainly distributed in the lens epithelial cells, with PP-1 as a major phosphatase. The mRNAs and proteins for PP-1cs and -2Acs are differentially expressed in the epithelial and fiber cells of rat and bovine lenses.

AB - PURPOSE. The reversible phosphorylation and dephosphorylation at the serine and threonine residues on proteins play distinct roles in regulating multiple cellular activities. Whereas the protein serine-threonine kinases have been well studied in the lens system, very little is known about the expression and function of the serine-threonine phosphatases. The present article reports the expression patterns of protein phosphatase (PP)-1 and -2A in adult rat and bovine lenses. METHODS. Total RNAs and proteins were extracted from the epithelial and fiber cells of rat and bovine lenses. RT-PCR and Northern blot analysis were used to detect the mRNA expression levels in the epithelial cells and different fractions of fiber cells of these two types of lenses. Western blot was used to examine the protein expression levels in these different samples. An enzymatic assay was used to detect the activity distribution of PP-1 and -2A in these samples. RESULTS. The mRNAs for the PP-1 catalytic subunit (PP-1cs) and PP-2A catalytic subunit (PP-2Acs) were expressed in both epithelial and fiber cells of rat and bovine lenses. A detailed examination of the expression patterns of the two mRNAs in different fractions of fiber cells revealed that the cortical fiber cells (F1) contain the highest level of PP-1cs and -2Acs mRNAs (similar to those in the epithelial cells) among different fractions of fiber cells. The levels of the two mRNAs were sequentially decreased in the next layers of fiber cells (F2 and F3) and became barely detectable in the inner layers of fiber cells (F4 and N). In contrast to the mRNA expression patterns, the PP-1cs protein was mainly found in the epithelial cells. Among different layers of fiber cells, only cortical (F1) fiber cells contained detectable level of PP-1cs protein (bovine lenses contained a relatively higher level of PP-1cs than rat lenses in this region). In the remaining fiber cells, the PP-1cs protein was hardly detectable in rat lenses and slightly detectable in bovine lenses. The PP-2Acs protein was detectable only in the lens epithelial cells. Enzymatic assays revealed that the distribution patterns of PP-1 and -2A activities were similar to those of PP-1cs and -2Acs proteins. Furthermore, PP-1 activity was approximately four to five times higher than PP-2A activity in the lens epithelial cells. CONCLUSIONS. This study demonstrates that active PP-1 and -2A are mainly distributed in the lens epithelial cells, with PP-1 as a major phosphatase. The mRNAs and proteins for PP-1cs and -2Acs are differentially expressed in the epithelial and fiber cells of rat and bovine lenses.

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