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Cloning and analysis of a novel conserved membrane zinc-metalloprotease family from Solanum surattense

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Abstract

Peptidases occur naturally in all organisms and their genes comprise 1–5% of the total number of genes. Genetic, biochemical, and molecular approaches used in recent years led to the identification and characterization of several plant organelle proteases, all of them being homologous to bacterial proteases best characterized in Escherichia coli. Here we report isolating and characterizing three novel genes, namely Sszn-mp1, Sszn-mp2, and SsZn-mp3 from Solanum surattense. To identify the subcellular location, structures, and functions of these three genes, integrated genomic approaches of data mining, expression profiling, and bioinformatic predictions were used. Sszn-mp is found to be constitutively expressed in tissues and regulated by various stimuli. Analysis of eight zinc-metalloproteases (Zn-MPs) deduced or assembled from Arabidopsis thaliana, tomato, potato, cotton, barley, sugarcane, and rice and four Zn-MPs from cyanobacteria (blue-green algae) in the GenBank database reveals that these proteins belong to a novel conserved membrane zinc-metalloprotease family. The plant Zn-MP members share more than 62% overall identity with SsZn-MP3, whereas four putative ATP-dependent zinc-proteases of cyanobacteria have low identity with SsZn-MP3 and their N-termini are about 110 amino acids shorter than those of plant Zn-MPs. The Zn-MP homologous sequences are found neither in other eukaryotic nor prokaryotic databases, suggesting that this family is specific to plants and cyanobacteria. The plant Zn-MP genes encoding membrane proteins are potentially targeted to chloroplast and plasma membranes, and the bacterial Zn-MPs are targeted to the cytoplasmic membrane, and their N-terminal targeting peptides are cleaved off for targeting the mature proteins to their subcellular compartments. The Zn-MP proteins contain a conserved zinc-binding site (HEAGHX19E/DX46∼48EX7E), a potential G-protein coupled receptors family 1 signature, and a triplet motif (N-R/K-F) in plant Zn-MPs, a D/E-R-Y motif in the four bacterial Zn-MPs, suggesting that the different mature forms of Zn-MPs may function as proteases and/or signal receptors.

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Abbreviations

aa:

amino acid

ER:

endoplasmic reticulum

EST:

expressed sequence tag

GPCR:

G-protein coupled receptor

GPI:

glycosylated phosphatidylinositol

MeJa:

methyl jasmonate

ORF:

open reading frame

PCR:

polymerase chain reaction

RT-PCR:

real time PCR

SA:

salicylic acid

TMD:

transmembrane domain

UTR:

untranslated region

Zn-MP:

zinc-metalloprotease

GrZn-MP:

Zn-MP from Gossipium raimondii

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Authors and Affiliations

  1. Shanghai Key Laboratory of Agrobiotechnology, Plant Biotechnology Research Center, School of Agriculture and Biology, School of Life Science and Technology, Fudan-SITU-Nottingham Plant Biotechnology R and D Center, Shanghai Jiao Tong University, Shanghai, 200030, China

    X. Sun & K. Tang

  2. State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai, 200433, China

    X. Sun, X. Liu, B. Huang, W. Li, X. Wang, J. Lin & K. Tang

  3. School of Life Sciences, Henan University, Kaifeng, 475001, China

    S. Guo

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  1. X. Sun
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  2. X. Liu
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  3. S. Guo
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  4. B. Huang
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Published in Russian in Fiziologiya Rastenii, 2007, Vol. 54, No. 1, pp. 73–84.

The text was submitted by the authors in English.

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Sun, X., Liu, X., Guo, S. et al. Cloning and analysis of a novel conserved membrane zinc-metalloprotease family from Solanum surattense . Russ J Plant Physiol 54, 63–73 (2007). https://doi.org/10.1134/S1021443707010104

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