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Characterization of SET/I2PP2A isoforms in dogs.

Yabe R, Fujiwara N, Mizuno T, Usui T, Ohama T, Sato K - J. Vet. Med. Sci. (2014)

Bottom Line: SET is an endogenous protein phosphatase 2A (PP2A) inhibitor and is associated with a poor prognosis in human leukemia.Here, we found that canine cells express several isoforms of the SET protein.We confirmed that SETα and β possess the ability to associate with PP2A.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Veterinary Pharmacology, Joint Faculty of Veterinary Medicine, Yamaguchi University, Yamaguchi 753-8515, Japan.

ABSTRACT
SET is an endogenous protein phosphatase 2A (PP2A) inhibitor and is associated with a poor prognosis in human leukemia. Previously, we reported increased SET protein levels in canine lymphoma cell lines and the potential therapeutic application of SET antagonists in canine lymphoma. Here, we found that canine cells express several isoforms of the SET protein. We cloned 4 isoforms of SET, named SETα, β, γ and δ. Genomic BLAST showed that the SET genes are located on chromosomes X, 7, 1 and 8, respectively. An immunofluorescent study showed nuclear localization of SETα and β, and nuclear and cytosolic localization of SETγ and δ. We confirmed that SETα and β possess the ability to associate with PP2A. Our data reveal the existence of unique SET isoforms that should be taken into account in SET-targeting drug development studies in dogs.

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Related in: MedlinePlus

SET protein expression in canine cells. SET protein expression in canine cells wasanalyzed by immunoblotting. Two types of anti-SET antibodies against 3–18 aa of humanSETα (N-term) and 175–215 aa of human SETα (Middle) were used. VCP was used as aloading control.
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fig_001: SET protein expression in canine cells. SET protein expression in canine cells wasanalyzed by immunoblotting. Two types of anti-SET antibodies against 3–18 aa of humanSETα (N-term) and 175–215 aa of human SETα (Middle) were used. VCP was used as aloading control.

Mentions: To examine the protein expression of SET isoforms in canine cells, normal peripheral bloodmononuclear cells (PBMC), melanoma cell lines CMeC1, CMeC2, KMeC and LMeC, lymphoma cellline Ema and the renal tubule epithelial cell line MDCK were lysed for immunoblotting. Themajor bands were observed around 37 kDa by 2 types of anti-SET antibodies against 3–18 aa ofhuman SETα (N-term) and 175–215 aa of human SETα (Middle). The minor bands were commonlyobserved around 25 kDa by the 2 antibodies. Only anti-SETα (Middle) detected the major bandaround 50 kDa and week bands around 15 kDa. These data indicate that canine cells expressmore than one SET isoform (Fig. 1Fig. 1.


Characterization of SET/I2PP2A isoforms in dogs.

Yabe R, Fujiwara N, Mizuno T, Usui T, Ohama T, Sato K - J. Vet. Med. Sci. (2014)

SET protein expression in canine cells. SET protein expression in canine cells wasanalyzed by immunoblotting. Two types of anti-SET antibodies against 3–18 aa of humanSETα (N-term) and 175–215 aa of human SETα (Middle) were used. VCP was used as aloading control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4197151&req=5

fig_001: SET protein expression in canine cells. SET protein expression in canine cells wasanalyzed by immunoblotting. Two types of anti-SET antibodies against 3–18 aa of humanSETα (N-term) and 175–215 aa of human SETα (Middle) were used. VCP was used as aloading control.
Mentions: To examine the protein expression of SET isoforms in canine cells, normal peripheral bloodmononuclear cells (PBMC), melanoma cell lines CMeC1, CMeC2, KMeC and LMeC, lymphoma cellline Ema and the renal tubule epithelial cell line MDCK were lysed for immunoblotting. Themajor bands were observed around 37 kDa by 2 types of anti-SET antibodies against 3–18 aa ofhuman SETα (N-term) and 175–215 aa of human SETα (Middle). The minor bands were commonlyobserved around 25 kDa by the 2 antibodies. Only anti-SETα (Middle) detected the major bandaround 50 kDa and week bands around 15 kDa. These data indicate that canine cells expressmore than one SET isoform (Fig. 1Fig. 1.

Bottom Line: SET is an endogenous protein phosphatase 2A (PP2A) inhibitor and is associated with a poor prognosis in human leukemia.Here, we found that canine cells express several isoforms of the SET protein.We confirmed that SETα and β possess the ability to associate with PP2A.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Veterinary Pharmacology, Joint Faculty of Veterinary Medicine, Yamaguchi University, Yamaguchi 753-8515, Japan.

ABSTRACT
SET is an endogenous protein phosphatase 2A (PP2A) inhibitor and is associated with a poor prognosis in human leukemia. Previously, we reported increased SET protein levels in canine lymphoma cell lines and the potential therapeutic application of SET antagonists in canine lymphoma. Here, we found that canine cells express several isoforms of the SET protein. We cloned 4 isoforms of SET, named SETα, β, γ and δ. Genomic BLAST showed that the SET genes are located on chromosomes X, 7, 1 and 8, respectively. An immunofluorescent study showed nuclear localization of SETα and β, and nuclear and cytosolic localization of SETγ and δ. We confirmed that SETα and β possess the ability to associate with PP2A. Our data reveal the existence of unique SET isoforms that should be taken into account in SET-targeting drug development studies in dogs.

Show MeSH
Related in: MedlinePlus