Characterization of Cucurbita maxima Phloem Serpin-1 (CmPS-1)

Yoo, Byung-Chun; Aoki, Koh; Yu, Xiang; Campbell, Leslie R.; Hull, Richard T.; Xoconostle‐Cázares, Beatriz; Monzer, Jan; Lee, Jinkee; Ullman, Diane E.; Lucas, William J.

doi:10.1074/jbc.m006060200

Cited by 82 publications

(52 citation statements)

References 47 publications

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“…Such an antipathogenic role could also be envisaged in potato, particularly as it is known that potato produces a range of different prophylactic defense proteins in tubers. Likewise, plants are known to have large amounts of functionally similar proteins in their phloem sap (Yoo et al, 2000 However, the expression of potato nsLTP genes indicates an apparently important role in new organ development, for example, during potato tuber life cycle. The expression profile of the three highly related TDFs displayed by cDNA-AFLP shows a short induction in the stolon just prior to tuber formation and again an increased expression at sprout development at the end of the dormancy period.…”

Section: Discussionmentioning

confidence: 99%

Expression Analysis of a Family ofnsLTPGenes Tissue Specifically Expressed throughout the Plant and during Potato Tuber Life Cycle

et al. 2002

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Non-specific lipid-transfer proteins (nsLTPs) are capable of binding lipid compounds in plant tissues and are coded by the nsLTP genes. Here, we present the analysis of expression of a family of potato (Solanum tuberosum) nsLTP genes that express throughout the developing plant in a highly tissue-specific manner. Three transcript-derived fragments were isolated using an amplified restriction fragment polymorphism-derived technique for RNA fingerprinting that show homology to plant nsLTP genes. These transcript-derived fragments displayed modulated expression profiles related to the development of new tissues, with a peak of transcription around the time of tuberization and just prior to sprout development, at dormancy breakage. In addition, a homologous family of expressed sequence tags was identified whose individual members could be classified according to their tissue specificity. Two subgroups of expressed sequence tags were found to express during tuber life cycle. To study the regulation of potato nsLTP genes, two putative potato nsLTP promoters were isolated and their expression was studied using promoter-marker-gene fusions. The results showed that one of the two promoters directed a highly specific pattern of expression detected in the phloem surrounding the nodes of young plants and in the same tissue of tuber related organs, whereas the second putative promoter showed little tissue or organ specificity. This difference in expression is likely due to a 331-bp insertion present in the tissue-specific promoter.The potato (Solanum tuberosum) tuber life cycle is a complex multistage process involving stolon formation, tuber initiation, tuber filling, dormancy, and sprouting (Cutter, 1978). Potato tubers constitute underground stems that have undergone a series of morphological changes. Depending on the environmental conditions and the genotype, the potato plant forms stolons at the base of the stem that, after a period of rapid longitudinal growth, swell at their tips initiating tuberization. The onset of tuberization is accompanied by a burst of cell division, and reorientation of the cytoskeleton occurs (Sanz et al., 1996). The morphological differentiation during tuberization is further characterized by a variety of biochemical changes, such as the accumulation of starch and the appearance of new proteins. At tuberization, suppression of growth is imposed on the axillary buds (eyes) of developing tubers leading into dormancy, at the end of which cell division is initiated in the eyes of the tuber, giving rise to sprout development.Each stage of tuber life cycle is likely to be controlled by a large set of interacting genes throughout the plant. Studies of gene expression using an amplified restriction fragment polymorphism-derived technique for RNA fingerprinting (cDNA-AFLP) during tuber life cycle show that many genes display differential expression and that these can be categorized into groups according to their putative function. The major processes that have been identified during tuber life cycle a...

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Section: Discussionmentioning

confidence: 99%

Expression Analysis of a Family ofnsLTPGenes Tissue Specifically Expressed throughout the Plant and during Potato Tuber Life Cycle

et al. 2002

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“…Interestingly, a correlation was found between a developmentally regulated increase in the amount of CmPS-1 and the reduced ability of the aphid, Myzus persicae, to survive. However, in that case, experiments with purified phloem CmPS-1 added to the aphid diet failed to demonstrate a direct effect on survival (17). Recently, an Arabidopsis serpin, AtSerpin1 (At1g47710), was reported to interact in vitro with the endogenous plant cysteine protease metacaspase 9 (AtMC9) (18).…”

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confidence: 99%

Arabidopsis AtSerpin1, Crystal Structure and in Vivo Interaction with Its Target Protease RESPONSIVE TO DESICCATION-21 (RD21)

Lampl

Budai-Hadrian

Davydov

et al. 2010

Journal of Biological Chemistry

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In animals, protease inhibitors of the serpin family are associated with many physiological processes, including blood coagulation and innate immunity. Serpins feature a reactive center loop (RCL), which displays a protease target sequence as a bait. RCL cleavage results in an irreversible, covalent serpin-protease complex. AtSerpin1 is an Arabidopsis protease inhibitor that is expressed ubiquitously throughout the plant. The x-ray crystal structure of recombinant AtSerpin1 in its native stressed conformation was determined at 2.2 Å . The electrostatic surface potential below the RCL was found to be highly positive, whereas the breach region critical for RCL insertion is an unusually open structure. AtSerpin1 accumulates in plants as a fulllength and a cleaved form. Fractionation of seedling extracts by nonreducing SDS-PAGE revealed the presence of an additional slower migrating complex that was absent when leaves were treated with the specific cysteine protease inhibitor L-trans-epoxysuccinyl-L-leucylamido (4-guanidino)butane. Significantly, RESPONSIVE TO DESICCATION-21 (RD21) was the major protease labeled with the L-trans-epoxysuccinyl-L-leucylamido (4-guanidino)butane derivative DCG-04 in wild type extracts but not in extracts of mutant plants constitutively overexpressing AtSerpin1, indicating competition. Fractionation by nonreducing SDS-PAGE followed by immunoblotting with RD21-specific antibody revealed that the protease accumulated both as a free enzyme and in a complex with AtSerpin1. Importantly, both RD21 and AtSerpin1 knock-out mutants lacked the serpin-protease complex. The results establish that the major Arabidopsis plant serpin interacts with RD21. This is the first report of the structure and in vivo interaction of a plant serpin with its target protease.Protease cascades are prominent mediators of rapid physiological responses in animals, playing a role in cellular immunity, blood clotting, and development. The proteolytic specificity of the serine and cysteine proteases involved dictates the fidelity of these reactions. The serpins are an important group of proteins that curb the activity of these cascades through specific irreversible inhibition of the proteases. For example, in Drosophila, the necrotic (nec) gene encodes a protease inhibitor of the serpin family. Necrotic protein controls a proteolytic cascade that activates the innate immune response to fungal and Gram-positive bacterial infections (1). In nec null mutants, Toll-mediated immune responses are constitutively activated, even in the absence of infection, implying that Nec continually restrains this immune response. As opposed to other types of protease inhibitors, serpins offer both an irreversible and tunable type of inhibition (reviewed in Ref.2). In their native conformation, serpins are in a stressed (spring-loaded) state with a solvent-exposed reactive center loop (RCL).3 Specific residues of the RCL are precisely accommodated by the target protease active site. Upon cleavage of the serpin peptide bond linking the P1 and ...

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“…The elastase cleavage sites within the reactive center loop of CmPS-1 were determined to be Val347-Gly348 and Val350-Ser351 with a 3:2 molar ratio. Importantly, in vivo feeding assays conducted with the aphid, Myzus persicae, established a close correlation between the developmentally regulated increase in CmPS-1 within the phloem sap and the reduced ability of these insects to survive and reproduce on pumpkin (Yoo et al 2000). These studies suggested that early expression of CmPS-1 might contribute to increased resistance to aphid infection.…”

Section: Characterization Of Phloem Serpin-1 (Cmps-1): a Developmentamentioning

confidence: 81%

FUNCTION OF PHLOEM-BORNE INFORMATION MACROMOLECULES IN INTEGRATING PLANT GROWTH & DEVELOPMENT

Lucas¹

2012

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Studies on higher plants have revealed the operation of cell-to-cell and long-distance communication networks that mediate the transport of information macromolecules, such as proteins and RNA. Based on the findings from this DOE-funded project and results from other groups, it is now well established that the enucleate sieve tube system of the angiosperms contains a complex set of proteins including RNA binding proteins as well as a unique population of RNA molecules, comprised of both mRNA and small RNA species. Heterografting experiments demonstrated that delivery of such RNA molecules, into the scion, is highly correlated with changes in developmental phenotypes. Furthermore, over the course of this project, our studies showed that plasmodesmata and the phloem are intimately involved in the local and systemic spread of sequence-specific signals that underlie gene silencing in plants. Major advances were also made in elucidating the underlying mechanisms that operate to mediate the selective entry and exit of proteins and RNA into and out of the phloem translocation stream. Our pioneering studies identified the first plant protein with the capacity to both bind specifically to small RNA molecules (si-RNA) and mediate in the cell-to-cell movement of such siRNA. Importantly, studies conducted with support from this DOE program also yielded a detailed characterization of the first phloem-mobile RNP complex isolated from pumpkin, namely the CmRBP50-RNP complex. This RNP complex was shown to bind, in a sequencespecific manner, to a set of transcripts encoding for transcription factors. The remarkable stability of this CmRBP50-RNP complex allows for long-distance delivery of bound transcripts from mature leaves into developing tissues and organs. Knowledge gained from this project can be used to exert control over the long-distance signaling networks used by plants to integrate their physiological and developmental programs at a whole plant level. Eventually, this information will aid in the engineering of elite plant lines with optimal traits for plant growth under non-ideal conditions, enhanced biomass and/or seed yield, and directed carbon allocation for efficient and sustainable biofuels production.

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Characterization of Cucurbita maxima Phloem Serpin-1 (CmPS-1)

Cited by 82 publications

References 47 publications

Expression Analysis of a Family ofnsLTPGenes Tissue Specifically Expressed throughout the Plant and during Potato Tuber Life Cycle

Expression Analysis of a Family ofnsLTPGenes Tissue Specifically Expressed throughout the Plant and during Potato Tuber Life Cycle

Arabidopsis AtSerpin1, Crystal Structure and in Vivo Interaction with Its Target Protease RESPONSIVE TO DESICCATION-21 (RD21)

FUNCTION OF PHLOEM-BORNE INFORMATION MACROMOLECULES IN INTEGRATING PLANT GROWTH & DEVELOPMENT

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