TY - JOUR
T1 - Soybean ATP sulfurylase, a homodimeric enzyme involved in sulfur assimilation, is abundantly expressed in roots and induced by cold treatment
AU - Phartiyal, Pallavi
AU - Kim, Won Seok
AU - Cahoon, Rebecca E.
AU - Jez, Joseph M.
AU - Krishnan, Hari B.
N1 - Funding Information:
The authors wish to thank Dr. Ahmed Mahmoud for Southern blot analysis, Dr. Joe Polacco (University of Missouri-Columbia) for providing the soybean seedling cDNA library, and Dr. Edgar Cahoon (Danforth Plant Science Center) for providing the Arabidopsis thaliana cDNA library. This work was funded in part by an U.S. Department of Agriculture grant (NRI-2005-02518) to J.M.J and an Illinois-Missouri Biotechnology Alliance grant to H.B.K. and J.M.J. Product names are necessary to report factually on available data; however, the University of Missouri, Danforth Plant Science Center and the USDA neither guarantees nor warrants the standard of product, and the use of the name by the University of Missouri and the USDA implies no approval of the product to the exclusion of others that may be suitable.
PY - 2006/6/1
Y1 - 2006/6/1
N2 - Soybeans are a rich source of protein and a key feed ingredient in livestock production, but lack sufficient levels of cysteine and methionine to meet the nutritional demands of swine or poultry as feed components. Although engineering the sulfur assimilatory pathway could lead to increased sulfur-containing amino acid content, little is known about this pathway in legumes. Here, we describe the cloning and characterization of soybean ATP sulfurylase (ATPS), which acts as the metabolic entry point into the sulfur assimilation pathway. Analysis of the ATPS clone isolated from a soybean seedling cDNA library revealed an open-reading frame, encoding a 52 kDa polypeptide with an N-terminal chloroplast/plastid transit peptide, which was related to the enzymes from Arabidopsis, potato, human, and yeast. Soybean ATP sulfurylase was expressed in Escherichia coli and purified to apparent homogeneity. Based on gel-filtration chromatography, the enzyme functions as a 100 kDa homodimer. Analysis of genomic DNA by Southern blotting revealed that multiple genes encode ATP sulfurylase in soybean. Analysis of the transcript profiles retrieved from a soybean EST database indicated that ATP sulfurylase mRNA was most abundant in root tissue. Cold treatment induced mRNA accumulation and enhanced the specific activity of ATP sulfurylase in root tissue. Northern blot analysis indicated a decline in the ATP sulfurylase transcript levels during seed development. Likewise, ATP sulfurylase specific activity also declined in the later stages of seed development. Increasing the expression levels of this key enzyme during soybean seed development could lead to an increase in the availability of sulfur amino acids, thereby enhancing the nutritional value of the crop.
AB - Soybeans are a rich source of protein and a key feed ingredient in livestock production, but lack sufficient levels of cysteine and methionine to meet the nutritional demands of swine or poultry as feed components. Although engineering the sulfur assimilatory pathway could lead to increased sulfur-containing amino acid content, little is known about this pathway in legumes. Here, we describe the cloning and characterization of soybean ATP sulfurylase (ATPS), which acts as the metabolic entry point into the sulfur assimilation pathway. Analysis of the ATPS clone isolated from a soybean seedling cDNA library revealed an open-reading frame, encoding a 52 kDa polypeptide with an N-terminal chloroplast/plastid transit peptide, which was related to the enzymes from Arabidopsis, potato, human, and yeast. Soybean ATP sulfurylase was expressed in Escherichia coli and purified to apparent homogeneity. Based on gel-filtration chromatography, the enzyme functions as a 100 kDa homodimer. Analysis of genomic DNA by Southern blotting revealed that multiple genes encode ATP sulfurylase in soybean. Analysis of the transcript profiles retrieved from a soybean EST database indicated that ATP sulfurylase mRNA was most abundant in root tissue. Cold treatment induced mRNA accumulation and enhanced the specific activity of ATP sulfurylase in root tissue. Northern blot analysis indicated a decline in the ATP sulfurylase transcript levels during seed development. Likewise, ATP sulfurylase specific activity also declined in the later stages of seed development. Increasing the expression levels of this key enzyme during soybean seed development could lead to an increase in the availability of sulfur amino acids, thereby enhancing the nutritional value of the crop.
KW - ATP sulfurylase
KW - Seed development
KW - Soybean
KW - Sulfur assimilation
UR - https://www.scopus.com/pages/publications/33646867610
U2 - 10.1016/j.abb.2006.03.033
DO - 10.1016/j.abb.2006.03.033
M3 - Article
C2 - 16684499
AN - SCOPUS:33646867610
SN - 0003-9861
VL - 450
SP - 20
EP - 29
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
IS - 1
ER -