Identification and characterization of a novel human plant pathogenesis-related protein that localizes to lipid-enriched microdomains in the Golgi complex

2002 ◽  
Vol 115 (4) ◽  
pp. 827-838 ◽  
Author(s):  
Heike B. Eberle ◽  
Ramon L. Serrano ◽  
Joachim Füllekrug ◽  
Andreas Schlosser ◽  
Wolf D. Lehmann ◽  
...  
Keyword(s):  
Immune System ◽  
Mammalian Cells ◽  
Electrostatic Interactions ◽  
Brefeldin A ◽  
Direct Interaction ◽  
Insoluble Fraction ◽  
Golgi Membranes ◽  
Pathogenesis Related Protein ◽  
Pathogenesis Related ◽  
Plant Pathogenesis

Group 1 of plant pathogenesis-related proteins (PR-1) and a variety of related mammalian proteins constitute a superfamily of proteins that share structural similarities. Little is known about their function, but all the family members identified to date are co-translationally translocated to the lumen of the endoplasmic reticulum and are secreted as soluble proteins or are targeted to vacuoles. Here we report the identification of a novel family member that localizes to the cytosolic site of the endomembrane system in mammalian cells. After detergent solubilization of isolated Golgi membranes, a 17 kDa protein was found associated with a low-density detergent-insoluble fraction. The amino-acid sequence, determined by microsequencing and molecular cloning, revealed a significant homology with the superfamily of PR-1 proteins. Golgi-associated PR-1 protein (GAPR-1) showed a brefeldin-A-sensitive Golgi localization in immunofluorescence. Interestingly,the protein remained associated with the microdomain fraction in the presence of Brefeldin A. By mass spectrometry, GAPR-1 was shown to be myristoylated. Immunoprecipitation of GAPR- 1 from Golgi membranes resulted in the coimmunoprecipitation of caveolin-1, indicating a direct interaction between these two proteins. Myristoylation, together with protein-protein or electrostatic interactions at physiological pH owing to the highly basic pI of GAPR-1 (pI 9.4) could explain the strong membrane association of GAPR-1. Tissue screening revealed that GAPR-1 is not detectably expressed in liver,heart or adrenal glands. High expression was found in monocytes, leukocytes,lung, spleen and embryonic tissue. Consistent with the involvement of PR-1 proteins in the plant immune system, these data could indicate that GAPR-1 is involved in the immune system.

scholarly journals Post-ER Stress Biogenesis of Golgi Is Governed by Giantin

Cells ◽  
2019 ◽  
Vol 8 (12) ◽  
pp. 1631
Author(s):  
Cole P. Frisbie ◽  
Alexander Y. Lushnikov ◽  
Alexey V. Krasnoslobodtsev ◽  
Jean-Jack M. Riethoven ◽  
Jennifer L. Clarke ◽  
...  
Keyword(s):  
Coiled Coil ◽  
Brefeldin A ◽  
Direct Interaction ◽  
Complete Recovery ◽  
Proximity Ligation Assay ◽  
Conformational Structure ◽  
Force Microscopy ◽  
Proximity Ligation ◽  
Golgi Membranes ◽  
Response To Stress

Background: The Golgi apparatus undergoes disorganization in response to stress, but it is able to restore compact and perinuclear structure under recovery. This self-organization mechanism is significant for cellular homeostasis, but remains mostly elusive, as does the role of giantin, the largest Golgi matrix dimeric protein. Methods: In HeLa and different prostate cancer cells, we used the model of cellular stress induced by Brefeldin A (BFA). The conformational structure of giantin was assessed by proximity ligation assay and atomic force microscopy. The post-BFA distribution of Golgi resident enzymes was examined by 3D SIM high-resolution microscopy. Results: We detected that giantin is rather flexible than an extended coiled-coil dimer and BFA-induced Golgi disassembly was associated with giantin monomerization. A fusion of the nascent Golgi membranes after BFA washout is forced by giantin re-dimerization via disulfide bond in its luminal domain and assisted by Rab6a GTPase. GM130-GRASP65-dependent enzymes are able to reach the nascent Golgi membranes, while giantin-sensitive enzymes appeared at the Golgi after its complete recovery via direct interaction of their cytoplasmic tail with N-terminus of giantin. Conclusion: Post-stress recovery of Golgi is conducted by giantin dimer and Golgi proteins refill membranes according to their docking affiliation rather than their intra-Golgi location.

scholarly journals BFR1, a multicopy suppressor of brefeldin A-induced lethality, is implicated in secretion and nuclear segregation in Saccharomyces cerevisiae.

Genetics ◽  
1994 ◽  
Vol 137 (2) ◽  
pp. 423-437 ◽  
Author(s):  
C L Jackson ◽  
F Képès
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mammalian Cells ◽  
Protein Transport ◽  
Brefeldin A ◽  
Secretory Proteins ◽  
Multicopy Suppressor ◽  
Dna Library ◽  
Golgi Membranes ◽  
Multicopy Suppressors ◽  
Genomic Dna Library

Abstract Brefeldin A (BFA) blocks protein transport out of the Golgi apparatus and causes disassembly of this organelle in mammalian cells. The primary effect of BFA is the release of the non-clathrin coat from Golgi membranes and vesicles. We sought to elucidate the mechanism of BFA action using a genetic approach in Saccharomyces cerevisiae. When an erg6 S. cerevisiae strain is treated with BFA, cell growth is arrested, cells lose viability and secretory proteins are accumulated in the endoplasmic reticulum (ER) and early Golgi compartments. We demonstrate that the mutant sec21 (defective in the S. cerevisiae homolog of gamma-COP, a non-clathrin coat protein) is supersensitive to BFA. Hence BFA probably affects the same processes in S. cerevisiae as in mammalian cells. We used a multicopy genomic DNA library to search for multicopy suppressors of BFA-induced lethality. We identified one such gene, BFR1, that, in addition, partially suppresses the growth and secretion defects of the ER-to-Golgi secretion mutant sec17. A bfr1-delta 1::URA3 deletion strain is viable, but has defects in cell morphology and nuclear segregation, and the mutation accentuates the growth and secretion defects of a sec21 mutant.

scholarly journals Current Overview of Allergens of Plant Pathogenesis Related Protein Families

2014 ◽  
Vol 2014 ◽  
pp. 1-19 ◽  
Author(s):  
Mau Sinha ◽  
Rashmi Prabha Singh ◽  
Gajraj Singh Kushwaha ◽  
Naseer Iqbal ◽  
Avinash Singh ◽  
...  
Keyword(s):  
Amino Acid ◽  
Cross Reactivity ◽  
Amino Acid Sequences ◽  
Pr Proteins ◽  
Antigenic Determinants ◽  
Related Protein ◽  
Protein Families ◽  
Pathogenesis Related Protein ◽  
Pathogenesis Related ◽  
Plant Pathogenesis

Pathogenesis related (PR) proteins are one of the major sources of plant derived allergens. These proteins are induced by the plants as a defense response system in stress conditions like microbial and insect infections, wounding, exposure to harsh chemicals, and atmospheric conditions. However, some plant tissues that are more exposed to environmental conditions like UV irradiation and insect or fungal attacks express these proteins constitutively. These proteins are mostly resistant to proteases and most of them show considerable stability at low pH. Many of these plant pathogenesis related proteins are found to act as food allergens, latex allergens, and pollen allergens. Proteins having similar amino acid sequences among the members of PR proteins may be responsible for cross-reactivity among allergens from diverse plants. This review analyzes the different pathogenesis related protein families that have been reported as allergens. Proteins of these families have been characterized in regard to their biological functions, amino acid sequence, and cross-reactivity. The three-dimensional structures of some of these allergens have also been evaluated to elucidate the antigenic determinants of these molecules and to explain the cross-reactivity among the various allergens.

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