scholarly journals A Family of Small Coiled-Coil–forming Proteins Functioning at the Late Endosome in Yeast

2001 ◽  
Vol 12 (3) ◽  
pp. 711-723 ◽  
Author(s):  
Andreas Kranz ◽  
Andrea Kinner ◽  
Ralf Kölling
Keyword(s):  
Sequence Analysis ◽  
Intracellular Trafficking ◽  
Coiled Coil ◽  
Open Reading Frame ◽  
Carboxypeptidase Y ◽  
Deletion Mutants ◽  
Differential Centrifugation ◽  
Reading Frame ◽  
Transporter Family ◽  
Vacuolar Protein

The multispanning membrane protein Ste6, a member of the ABC-transporter family, is transported to the yeast vacuole for degradation. To identify functions involved in the intracellular trafficking of polytopic membrane proteins, we looked for functions that block Ste6 transport to the vacuole upon overproduction. In our screen, we identified several known vacuolar protein sorting (VPS) genes (SNF7/VPS32,VPS4, and VPS35) and a previously uncharacterized open reading frame, which we named MOS10(more of Ste6). Sequence analysis showed that Mos10 is a member of a small family of coiled-coil–forming proteins, which includes Snf7 and Vps20. Deletion mutants of all three genes stabilize Ste6 and show a “class E vps phenotype.” Maturation of the vacuolar hydrolase carboxypeptidase Y was affected in the mutants and the endocytic tracer FM4-64 and Ste6 accumulated in a dot or ring-like structure next to the vacuole. Differential centrifugation experiments demonstrated that about half of the hydrophilic proteins Mos10 and Vps20 was membrane associated. The intracellular distribution was further analyzed for Mos10. On sucrose gradients, membrane-associated Mos10 cofractionated with the endosomal t-SNARE Pep12, pointing to an endosomal localization of Mos10. The growth phenotypes of the mutants suggest that the “Snf7-family” members are involved in a cargo-specific event.

scholarly journals Open reading frame 132 of Heliocoverpa armigera nucleopolyhedrovirus encodes a functional per os infectivity factor (PIF-2)

2006 ◽  
Vol 87 (9) ◽  
pp. 2563-2569 ◽  
Author(s):  
Minggang Fang ◽  
Yingchao Nie ◽  
Qian Wang ◽  
Fei Deng ◽  
Ranran Wang ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Western Blot ◽  
Blot Analysis ◽  
Post Infection ◽  
Spodoptera Exigua ◽  
Open Reading Frame ◽  
Rt Pcr ◽  
Western Blots ◽  
Reading Frame ◽  
Budded Virus

Open reading frame 132 (Ha132) of Helicoverpa armigera nucleopolyhedrovirus (HearNPV) is a homologue of per os infectivity factor 2 (pif-2) of Spodoptera exigua multiple nucleopolyhedrovirus. Sequence analysis indicated that Ha132 encoded a protein of 383 aa with a predicted molecular mass of 44.5 kDa. Alignment of HA132 and its baculovirus homologues revealed that HA132 was highly conserved among baculoviruses, with 14 absolutely conserved cysteine residues. RT-PCR indicated that Ha132 was first transcribed at 24 h post-infection. Western blot analysis showed that a 43 kDa band was detectable in HearNPV-infected HzAM1 cells from 36 h post-infection. Western blots also indicated that HA132 was a component of the occlusion-derived virus, but not of budded virus. Deletion of Ha132 from HearNPV abolished per os infectivity, but had no effect on the infectivity of the budded virus phenotype.

Sequence of a Rhodococcus gene cluster encoding the subunits of ethanolamine ammonia-lyase and an APC-like permease

1994 ◽  
Vol 40 (5) ◽  
pp. 403-407 ◽  
Author(s):  
René De Mot ◽  
Istvan Nagy ◽  
Geert Schoofs ◽  
Jos Vanderleyden
Keyword(s):  
Open Reading Frame ◽  
Genomic Fragment ◽  
Positive Bacterium ◽  
Gram Positive ◽  
Gene Encoding ◽  
Reading Frame ◽  
Transporter Family ◽  
Rhodococcus Species ◽  
Rhodococcus Sp

Sequence analysis of a 5173-bp genomic fragment from the nocardioform actinomycete Rhodococcus sp. strain NI86/21 revealed the presence of two genes, eutB and eutC, encoding the putative homologues of the large and small subunits of the ethanolamine ammonia-lyase, respectively, from Salmonella typhimurium. This is the first report of the characterization of these genes in a Gram-positive species. Immediately upstream of eutB, a gene encoding a putative permease of the APC (amino acids, polyamines, choline) transporter family was located. At present, no other Gram-positive members of this permease family are known. The translational coupling of these eut genes suggests an operon-like organization of the ethanolamine genes in Rhodococcus species. A truncated open reading frame downstream of eutC contained an N-terminal motif characteristic of membrane-anchored lipoproteins.Key words: nocardioform actinomycete, cobalamin, APC transporter, membrane-anchored lipoprotein, Gram-positive bacterium.

scholarly journals Outbreak of equid herpesvirus 1 abortions at the Arabian stud in Poland

2020 ◽  
Vol 16 (1) ◽  
Author(s):  
Karol Stasiak ◽  
Magdalena Dunowska ◽  
Jerzy Rola
Keyword(s):  
Amino Acid ◽  
Sequence Analysis ◽  
Amino Acid Sequence ◽  
Neurological Disease ◽  
Open Reading Frame ◽  
Case Presentation ◽  
Reading Frame ◽  
Equid Herpesvirus ◽  
Herpesvirus 1 ◽  
Horizontal Spread

Abstract Background Equid herpesvirus 1 (EHV-1) infections are endemic worldwide, including Poland. Many are subclinical, but some are associated with respiratory disease, abortion, neonatal foal death, or neurological disease. We describe an outbreak of abortions in Arabian mares at a well-managed State stud farm in Poland. Case presentation Eight of 30 pregnant mares aborted and one gave birth to a weak foal that died within 72 h after birth. EHV-1 was isolated from all fetuses as well as from the diseased foal. All viruses belonged to the N752 variant based on the predicted open reading frame (ORF) 30 amino acid sequence. All were identical to each other and to previous EHV-1 viruses from the same stud based on the ORF68 sequence analysis. The outbreak coincided with the lapse in the routine yearly EHV-1/4 vaccinations of the mares. Conclusions Multiple abortion due to EHV-1 infection can occur in well-managed groups of horses. Reactivation of latent EHV-1 in one of the resident mares followed by a horizontal spread was considered the most likely explanation for the outbreak. Routine vaccination is an important part of a herd-heath program.

scholarly journals Structural and Functional Analysis of a Novel Coiled-Coil Protein Involved in Ypt6 GTPase-regulated Protein Transport in Yeast

1999 ◽  
Vol 10 (1) ◽  
pp. 63-75 ◽  
Author(s):  
Miki Tsukada ◽  
Elke Will ◽  
Dieter Gallwitz
Keyword(s):  
Protein Transport ◽  
Protein Complexes ◽  
Coiled Coil ◽  
Carboxypeptidase Y ◽  
Loss Of Function ◽  
Golgi Compartment ◽  
Helical Content ◽  
Associated Proteins ◽  
Yeast Genes ◽  
Vacuolar Protein

The yeast transport GTPase Ypt6p is dispensable for cell growth and secretion, but its lack results in temperature sensitivity and missorting of vacuolar carboxypeptidase Y. We previously identified four yeast genes (SYS1, 2, 3, and 5) that on high expression suppressed these phenotypic alterations.SYS3 encodes a 105-kDa protein with a predicted high α-helical content. It is related to a variety of mammalian Golgi-associated proteins and to the yeast Uso1p, an essential protein involved in docking of endoplasmic reticulum–derived vesicles to thecis-Golgi. Like Uso1p, Sys3p is predominatly cytosolic. According to gel chromatographic, two-hybrid, and chemical cross-linking analyses, Sys3p forms dimers and larger protein complexes. Its loss of function results in partial missorting of carboxypeptidase Y. Double disruptions of SYS3and YPT6 lead to a significant growth inhibition of the mutant cells, to a massive accumulation of 40- to 50-nm vesicles, to an aggravation of vacuolar protein missorting, and to a defect in α-pheromone processing apparently attributable to a perturbation of protease Kex2p cycling between the Golgi and a post-Golgi compartment. The results of this study suggest that Sys3p, like Ypt6p, acts in vesicular transport (presumably at a vesicle-docking stage) between an endosomal compartment and the most distal Golgi compartment.

Hepatitis B X open reading frame deletion mutants isolated from atypical hepatitis B virus infections

1991 ◽  
Vol 13 ◽  
pp. S58-S60 ◽  
Author(s):  
Mark Feitelson ◽  
Ling-Xun Duan ◽  
Norio Horiike ◽  
Marcia Clayton
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Open Reading Frame ◽  
Deletion Mutants ◽  
Virus Infections ◽  
Reading Frame ◽  
B Virus ◽  
Hepatitis B Virus Infections

scholarly journals Glycine Betaine Transmethylase Mutant of Pseudomonas aeruginosa

2002 ◽  
Vol 184 (15) ◽  
pp. 4301-4303 ◽  
Author(s):  
Ana L. Serra ◽  
Javier F. Mariscotti ◽  
José L. Barra ◽  
Gloria I. Lucchesi ◽  
Carlos E. Domenech ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Sequence Analysis ◽  
Glycine Betaine ◽  
Nitrogen Sources ◽  
Open Reading Frame ◽  
Carbon And Nitrogen Sources ◽  
Reading Frame ◽  
Carbon And Nitrogen ◽  
Molecular Approaches ◽  
Knockout Gene

ABSTRACT The gene for glycine betaine transmethylase (gbt) was identified in Pseudomonas aeruginosa strain Fildes III by biochemical, physiological, and molecular approaches. Based on sequence analysis, the knockout gene corresponded to an open reading frame (ORF) named PA3082 in the genome of P. aeruginosa PAO1. The translated product of this ORF displayed similarity to transferases of different microorganisms. Mutation in gbt blocked the utilization of choline and glycine betaine as carbon and nitrogen sources.

scholarly journals Knot Formation Caused by Pseudomonas syringae subsp. savastanoi on Olive Plants Is hrp-Dependent

2004 ◽  
Vol 94 (5) ◽  
pp. 484-489 ◽  
Author(s):  
A. Sisto ◽  
M. G. Cipriani ◽  
M. Morea
Keyword(s):  
Sequence Analysis ◽  
Pseudomonas Syringae ◽  
Open Reading Frame ◽  
Gene Encoding ◽  
Disease Symptoms ◽  
Phytopathogenic Bacteria ◽  
Reading Frame ◽  
Nonhost Plant ◽  
Induced Mutant

The virulence of Pseudomonas syringae subsp. savastanoi, which causes hyperplastic symptoms (knots) on olive plants, is associated with secreted phytohormones. We identified a Tn5-induced mutant of P. syringae subsp. savastanoi that did not cause disease symptoms on olive plants although it was still able to produce phytohormones. In addition, the mutant failed to elicit a hypersensitive response in a nonhost plant. Molecular characterization of the mutant revealed that a single Tn5 insertion occurred within an open reading frame encoding a protein 92% identical to the HrcC protein of P. syringae pv. syringae. Moreover, sequence analysis revealed that the gene encoding the HrcC protein in P. syringae subsp. savastanoi was part of an operon that included five genes arranged as in other phytopathogenic bacteria. These results imply that hrp/hrc genes are functional in P. syringae subsp. savastanoi and that they play a key role in the pathogenicity of this plant pathogen.

scholarly journals Luv1p/Rki1p/Tcs3p/Vps54p, a Yeast Protein That Localizes to the Late Golgi and Early Endosome, Is Required for Normal Vacuolar Morphology.

2000 ◽  
Vol 11 (7) ◽  
pp. 2429-2443 ◽  
Author(s):  
Michael J. Conboy ◽  
Martha S. Cyert
Keyword(s):  
Fluorescent Protein ◽  
Protein A ◽  
Coiled Coil ◽  
Specific Inhibitor ◽  
Early Endosome ◽  
Carboxypeptidase Y ◽  
Similar Drug ◽  
Green Fluorescent ◽  
Vacuolar Protein ◽  
Golgi Network

We have characterized LUV1/RKI1/TCS3/VPS54, a novel yeast gene required to maintain normal vacuolar morphology. Theluv1 mutant was identified in a genetic screen for mutants requiring the phosphatase calcineurin for vegetative growth.luv1 mutants lack a morphologically intact vacuole and instead accumulate small vesicles that are acidified and contain the vacuolar proteins alkaline phosphatase and carboxypeptidase Y and the vacuolar membrane H+-ATPase. Endocytosis appears qualitatively normal in luv1 mutants, but some portion (28%) of carboxypeptidase Y is secreted. luv1 mutants are sensitive to several ions (Zn2+, Mn2+, and Cd2+) and to pH extremes. These mutants are also sensitive to hygromycin B, caffeine, and FK506, a specific inhibitor of calcineurin. Some vacuolar protein-sorting mutants display similar drug and ion sensitivities, including sensitivity to FK506. Luv1p sediments at 100,000 × g and can be solubilized by salt or carbonate, indicating that it is a peripheral membrane protein. A Green Fluorescent Protein–Luv1 fusion protein colocalizes with the dye FM 4-64 at the endosome, and hemagglutinin-tagged Luv1p colocalizes with the trans-Golgi network/endosomal protease Kex2p. Computer analysis predicts a short coiled-coil domain in Luv1p. We propose that this protein maintains traffic through or the integrity of the early endosome and that this function is required for proper vacuolar morphology.

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