Molecular Cloning of the Genes Encoding the PR55/Bβ/δ Regulatory Subunits for PP-2A and Analysis of Their Functions in Regulating Development of Goldfish, Carassius auratus.
The protein phosphatase-2A (PP-2A), one of the major phosphatases in eukaryotes, is a heterotrimer, consisting of a scaffold A subunit, a catalytic C subunit and a regulatory B subunit. Previous studies have shown that besides regulating specific PP-2A activity, various B subunits encoded by more than 16 different genes, may have other functions. To explore the possible roles of the regulatory subunits of PP-2A in vertebrate development, we have cloned the PR55/B family regulatory subunits: β and δ, analyzed their tissue specific and developmental expression patterns in Goldfish ( Carassius auratus). Our results revealed that the full-length cDNA for PR55/Bβ consists of 1940 bp with an open reading frame of 1332 nucleotides coding for a deduced protein of 443 amino acids. The full length PR55/Bδ cDNA is 2163 bp containing an open reading frame of 1347 nucleotides encoding a deduced protein of 448 amino acids. The two isoforms of PR55/B display high levels of sequence identity with their counterparts in other species. The PR55/Bβ mRNA and protein are detected in brain and heart. In contrast, the PR55/Bδ is expressed in all 9 tissues examined at both mRNA and protein levels. During development of goldfish, the mRNAs for PR55/Bβ and PR55/Bδ show distinct patterns. At the protein level, PR55/Bδ is expressed at all developmental stages examined, suggesting its important role in regulating goldfish development. Expression of the PR55/Bδ anti-sense RNA leads to significant downregulation of PR55/Bδ proteins and caused severe abnormality in goldfish trunk and eye development. Together, our results suggested that PR55/Bδ plays an important role in governing normal trunk and eye formation during goldfish development.
Key Laboratory of Protein Chemistry and Developmental Biology of Educational Ministry of China, College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China.
Jun-Qiong
JQ
Zhao
Si-Si
SS
Xie
Wen-Bin
WB
Liu
Ya-Mei
YM
Xiao
Xiao-Ming
XM
Zeng
Mi
M
Deng
Lili
L
Gong
Jin-Ping
JP
Liu
Pei-Chao
PC
Chen
Jie
J
Zhou
Xiao-Hui
XH
Hu
Jia-Han
JH
Lv
Xiang-Qian
XQ
Yu
Dao
D
Wang
Chi
C
Li
Yun-Lei
YL
Peng
Gao-Peng
GP
Liao
Yun
Y
Liu
David Wan-Cheng
DW
Li
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