Expression patterns of purple acid phosphatase genes in Arabidopsis organs and functional analysis of AtPAP23 predominantly transcribed in flower.
Purple acid phosphatases (PAPs) are metallo-phosphoesterases. Their expression and function have not been systematically investigated in higher plants. In this work, we compared the transcript levels of 28 Arabidopsis PAP (AtPAP) genes in five Arabidopsis organs. The 28 members, although differed in their expression patterns in vegetative organs, were all transcribed in flower. Furthermore, the transcription of seven members (AtPAPs 6, 11, 14, 19, 23, 24 and 25) occurred predominantly in the flower. To begin dissecting the role of AtPAP genes in flower development, further expression and functional analyses were conducted using AtPAP23. Histochemical staining of transgenic plants expressing AtPAP23 promoter-beta-glucuronidase (GUS) gene construct revealed that AtPAP23 transcription was strong in flower apical meristems, but became restricted to petals and anther filaments in fully developed flower. A GST (glutathione S-transferase) fusion protein of AtPAP23 (GST:AtPAP23) was expressed in bacterial cells, and was found to contain significant amounts of Fe and Mn (whereas the control GST protein contained none). In biochemical tests, GST:AtPAP23 showed typical acid phosphatase activities. The fusion protein was also highly active on phosphoserine, but not phosphotyrosine. Despite its highly specific expression pattern and the demonstrated biochemical function of its protein product, the RNAi (RNA interference), T-DNA knock-out and overexpression lines of AtPAP23 were indistinguishable from wild type plants in the development of flower (or other organs). Interestingly, the Fe and Mn contents were found significantly increased in AtPAP23 overexpression lines, which may offer a new direction for further functional studies of AtPAPs in Arabidopsis.
Zhu
Huifen
H
Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Datun Road, 100101 Beijing, Chaoyang District, China.
Qian
Weiqiang
W
Lu
Xuzhong
X
Li
Dongping
D
Liu
Xin
X
Liu
Kunfan
K
Wang
Daowen
D
D000135:Acid Phosphatase / Q000235:genetics* / Q000378:metabolism*; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics*; D029681:Arabidopsis Proteins; D003001:Cloning, Molecular; D035264:Flowers / Q000201:enzymology / Q000235:genetics*; D017353:Gene Deletion; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D006023:Glycoproteins / Q000235:genetics* / Q000378:metabolism*; D007501:Iron / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D009097:Multienzyme Complexes; D034622:RNA Interference; D014158:Transcription, Genetic / Q000235:genetics
D029681:Arabidopsis Proteins; D006023:Glycoproteins; D009097:Multienzyme Complexes; D008345:Manganese; D007501:Iron; C529220:PAP15 protein, Arabidopsis; C091137:purple acid phosphatase; D000135:Acid Phosphatase; C506758:PAP23 protein, Arabidopsis