scholarly journals Functional Characterization of Pneumocystis carinii brl1 by Transspecies Complementation Analysis

2007 ◽  
Vol 6 (12) ◽  
pp. 2448-2452 ◽  
Author(s):  
Libera Lo Presti ◽  
Moira Cockell ◽  
Lorenzo Cerutti ◽  
Viesturs Simanis ◽  
Philippe M. Hauser
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Membrane Proteins ◽  
Nuclear Membrane ◽  
Opportunistic Infections ◽  
Functional Characterization ◽  
Pneumocystis Carinii ◽  
Ectopic Expression ◽  
Functional Complementation ◽  
Null Alleles

ABSTRACT Pneumocystis jirovecii is a fungus which causes severe opportunistic infections in immunocompromised humans. The brl1 gene of P. carinii infecting rats was identified and characterized by using bioinformatics in conjunction with functional complementation in Saccharomyces cerevisiae and Schizosaccharomyces pombe. The ectopic expression of this gene rescues null alleles of essential nuclear membrane proteins of the Brr6/Brl1 family in both yeasts.

Novel biochemical and functional insights into nuclear Ca2+ transport through IP3Rs and RyRs in osteoblasts

2000 ◽  
Vol 278 (5) ◽  
pp. F784-F791 ◽  
Author(s):  
Olugbenga A. Adebanjo ◽  
Gopa Biswas ◽  
Baljit S. Moonga ◽  
Hindupur K. Anandatheerthavarada ◽  
Li Sun ◽  
...  
Keyword(s):  
Gene Expression ◽  
Nuclear Envelope ◽  
Nuclear Membrane ◽  
Ryanodine Receptors ◽  
Functional Characterization ◽  
Inositol Trisphosphate Receptors ◽  
Nuclear Membranes ◽  
Sds Page ◽  
Direct Regulation

We report the first biochemical and functional characterization of inositol trisphosphate receptors (IP3Rs) and ryanodine receptors (RyRs) in the nuclear membrane of bone-forming (MC3T3-E1) osteoblasts. Intact nuclei fluoresced intensely with anti-RyR (Ab34) and anti-IP3R (Ab40) antisera in a typically peripheral nuclear membrane pattern. Isolated nuclear membranes were next subjected to SDS-PAGE and blotted with isoform-specific anti-receptor antisera, notably Ab40, anti-RyR-1, anti-RyR-2 (Ab129), and anti-RyR-3 (Ab180). Only anti-RyR-1 and Ab40 showed bands corresponding, respectively, to full-length RyR-1 (∼500 kDa) and IP3R-1 (∼250 kDa). Band intensity was reduced by just ∼20% after brief tryptic proteolysis of intact nuclei; this confirmed that isolated nuclear membranes were mostly free of endoplasmic reticular contaminants. Finally, the nucleoplasmic Ca2+ concentration ([Ca2+]np) was measured in single nuclei by using fura-dextran. The nuclear envelope was initially loaded with Ca2+ via Ca2+-ATPase activation (1 mM ATP and ∼100 nM Ca2+). Adequate Ca2+ loading was next confirmed by imaging the nuclear envelope (and nucleoplasm). Exposure of Ca2+-loaded nuclei to IP3 or cADP ribose resulted in a rapid and sustained [Ca2+]np elevation. Taken together, the results provide complementary evidence for nucleoplasmic Ca2+ influx in osteoblasts through nuclear membrane-resident IP3Rs and RyRs. Our findings may conceivably explain the direct regulation of osteoblastic gene expression by hormones that use the IP3-Ca2+pathway.

scholarly journals Structural Characterization of the LEM Motif Common to Three Human Inner Nuclear Membrane Proteins

Structure ◽  
2001 ◽  
Vol 9 (6) ◽  
pp. 503-511 ◽  
Author(s):  
Cédric Laguri ◽  
Bernard Gilquin ◽  
Nicolas Wolff ◽  
Régine Romi-Lebrun ◽  
Karine Courchay ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Structural Characterization ◽  
Nuclear Membrane ◽  
Inner Nuclear Membrane

scholarly journals Molecular Characterization of the Interaction of Borrelia parkeri and Borrelia turicatae with Human Complement Regulators

2010 ◽  
Vol 78 (5) ◽  
pp. 2199-2208 ◽  
Author(s):  
Melanie Schott ◽  
Sonja Grosskinsky ◽  
Christiane Brenner ◽  
Peter Kraiczy ◽  
Reinhard Wallich
Keyword(s):  
Binding Capacity ◽  
Functional Characterization ◽  
Ectopic Expression ◽  
Factor H ◽  
Human Complement ◽  
Related Protein ◽  
Relapsing Fever ◽  
Extracellular Matrix Components ◽  
Complement Regulators

ABSTRACT In North America, tick-borne relapsing fever is caused by the species Borrelia hermsii, B. parkeri, and B. turicatae, which are transmitted to humans through the bite of the respective infected tick vectors. Here we describe the identification and functional characterization of a surface lipoprotein of B. parkeri, designated BpcA, that binds the human complement regulators factor H and factor H-related protein 1 and, simultaneously, the host protease plasminogen. In contrast, the homologous B. turicatae protein failed to bind human factor H and factor H-related protein 1 but retained its plasminogen binding capacity. Factor H bound to BpcA maintains its regulatory capacity to control C3b deposition and C3 convertase activity. Ectopic expression of BpcA in a serum-sensitive B. burgdorferi strain protects transformed cells from complement-mediated killing. Furthermore, bound plasminogen/plasmin endows B. parkeri and B. turicatae with the potential to degrade extracellular matrix components. These findings expand our understanding of the putative recent evolutionary separation of Borrelia parkeri and Borrelia turicatae, provide evidence that B. parkeri differs from B. turicatae in its ability to resist complement attack, and may help in understanding the pathological processes underlying tick-borne relapsing fever.

Isolation and functional characterization of mutant ferrochelatases in Saccharomyces cerevisiae

Biochimie ◽  
1996 ◽  
Vol 78 (2) ◽  
pp. 144-152 ◽  
Author(s):  
M. Góra ◽  
A. Chaciñska ◽  
J. Rytka ◽  
R. Labbe-Bois
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Functional Characterization

Use of Monoclonal Antibodies in the Functional Characterization of the Saccharomyces Cerevisiae Sepl Protein

1995 ◽  
Vol 231 (2) ◽  
pp. 329-336 ◽  
Author(s):  
Anita Holler ◽  
Vladimir I. Bashkirov ◽  
Jachen A. Solinger ◽  
Ursula Reinhart ◽  
Wolf-Dietrich Heyer
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Monoclonal Antibodies ◽  
Functional Characterization

scholarly journals Functional Characterization of BRASSINAZOLE-RESISTANT 1 in Panax Ginseng (PgBZR1) and Brassinosteroid Response during Storage Root Formation

2020 ◽  
Vol 21 (24) ◽  
pp. 9666
Author(s):  
Hyeona Hwang ◽  
Hwa-Yong Lee ◽  
Hojin Ryu ◽  
Hyunwoo Cho
Keyword(s):  
Panax Ginseng ◽  
Glycogen Synthase ◽  
Functional Characterization ◽  
Secondary Growth ◽  
Ectopic Expression ◽  
Floral Organ ◽  
Membrane Receptors ◽  
Storage Root ◽  
Functional Conservation

Brassinosteroids (BRs) play crucial roles in the physiology and development of plants. In the model plant Arabidopsis, BR signaling is initiated at the level of membrane receptors, BRASSINOSTEROIDS INSENSITIVE 1 (BRI1) and BRI1-ASSOCIATED RECEPTOR KINASE 1 (BAK1) complex, thus activating the transcription factors (TFs) BRASSINAZOLE RESISTANT 1/BRI1-EMS-SUPPRESSOR 1 (BZR1/BES1) to coordinate BR responsive genes. BRASSINOSTEROIDS INSENSITIVE 2 (BIN2), glycogen synthase kinase 3 (GSK3) like-kinase, negatively regulates BZR1/BES1 transcriptional activity through phosphorylation-dependent cytosolic retention and shuttling. However, it is still unknown whether this mechanism is conserved in Panax ginseng C. A. Mayer, a member of the Araliaceae family, which is a shade-tolerant perennial root crop. Despite its pharmacological and agricultural importance, the role of BR signaling in the development of P. ginseng and characterization of BR signaling components are still elusive. In this study, by utilizing the Arabidopsisbri1 mutant, we found that ectopic expression of the gain of function form of PgBZR1 (Pgbzr1-1D) restores BR deficiency. In detail, ectopic expression of Pgbzr1-1D rescues dwarfism, defects of floral organ development, and hypocotyl elongation of bri1-5, implying the functional conservation of PgBZR1 in P. ginseng. Interestingly, brassinolide (BL) and BRs biosynthesis inhibitor treatment in two-year-old P. ginseng storage root interferes with and promotes, respectively, secondary growth in terms of xylem formation. Altogether, our results provide new insight into the functional conservation and potential diversification of BR signaling and response in P. ginseng.

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