scholarly journals Isolation and characterization of yeast mutants in the cytoplasm to vacuole protein targeting pathway.

1995 ◽  
Vol 131 (3) ◽  
pp. 591-602 ◽  
Author(s):  
T M Harding ◽  
K A Morano ◽  
S V Scott ◽  
D J Klionsky
Keyword(s):  
Secretory Pathway ◽  
Protein Targeting ◽  
Protein Localization ◽  
Amino Terminal ◽  
Isolation And Characterization ◽  
Alternate Mechanism ◽  
Complementation Groups ◽  
Vacuolar Protein ◽  
Yeast Mutants

In Saccharomyces cerevisiae the vacuolar protein aminopeptidase I (API) is localized to the vacuole independent of the secretory pathway. The alternate targeting mechanism used by this protein has not been characterized. API is synthesized as a 61-kD soluble cytosolic precursor. Upon delivery to the vacuole, the amino-terminal propeptide is removed by proteinase B (PrB) to yield the mature 50-kD hydrolase. We exploited this delivery-dependent maturation event in a mutant screen to identify genes whose products are involved in API targeting. Using antiserum to the API propeptide, we isolated mutants that accumulate precursor API. These mutants, designated cvt, fall into eight complementation groups, five of which define novel genes. These five complementation groups exhibit a specific defect in maturation of API, but do not have a significant effect on vacuolar protein targeting through the secretory pathway. Localization studies show that precursor API accumulates outside of the vacuole in all five groups, indicating that they are blocked in API targeting and/or translocation. Future analysis of these gene products will provide information about the subcellular components involved in this alternate mechanism of vacuolar protein localization.

scholarly journals New Mutants of Saccharomyces cerevisiae Affected in the Transport of Proteins From the Endoplasmic Reticulum to the Golgi Complex

Genetics ◽  
1996 ◽  
Vol 142 (2) ◽  
pp. 393-406 ◽  
Author(s):  
Linda J Wuestehube ◽  
Rainer Duden ◽  
Arlene Eun ◽  
Susan Hamamoto ◽  
Paul Korn ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Complex ◽  
Secretory Pathway ◽  
Secretory Protein ◽  
Temperature Sensitive ◽  
Nonpermissive Temperature ◽  
Membrane Structures ◽  
Α Subunit ◽  
Complementation Groups ◽  
Vacuolar Protein

Abstract We have isolated new temperature-sensitive mutations in five complementation groups, sec31-sec35, that are defective in the transport of proteins from the endoplasmic reticulum (ER) to the Golgi complex. The sec31-sec35 mutants and additional alleles of previously identified sec and vacuolar protein sorting (vps) genes were isolated in a screen based on the detection of α-factor precursor in yeast colonies replicated to and lysed on nitrocellulose filters. Secretory protein precursors accumulated in sec31-sec35 mutants at the nonpermissive temperature were core-glycosylated but lacked outer chain carbohydrate, indicating that transport was blocked after translocation into the ER but before arrival in the Golgi complex. Electron microscopy revealed that the newly identified sec mutants accumulated vesicles and membrane structures reminiscent of secretory pathway organelles. Complementation analysis revealed that sec32-1 is an allele of BOS1, a gene implicated in vesicle targeting to the Golgi complex, and sec33-1 is an allele of RET1, a gene that encodes the α subunit of coatomer.

scholarly journals Isolation and characterization of awamori yeast mutants with l-leucine accumulation that overproduce isoamyl alcohol

2015 ◽  
Vol 119 (2) ◽  
pp. 140-147 ◽  
Author(s):  
Hiroshi Takagi ◽  
Keisuke Hashida ◽  
Daisuke Watanabe ◽  
Ryo Nasuno ◽  
Masataka Ohashi ◽  
...  
Keyword(s):  
Isoamyl Alcohol ◽  
Isolation And Characterization ◽  
Yeast Mutants

Isolation and characterization of yeast mutants blocked in mevalonic acid formation

1984 ◽  
Vol 123 (2) ◽  
pp. 424-430 ◽  
Author(s):  
Madeleine Servouse ◽  
Nicole Mons ◽  
Jean-Louis Baillargeat ◽  
Francis Karst
Keyword(s):  
Mevalonic Acid ◽  
Acid Formation ◽  
Isolation And Characterization ◽  
Yeast Mutants

Isolation and characterization of human pro-urokinase and its mutants accumulated within the yeast secretory pathway

Gene ◽  
1991 ◽  
Vol 99 (2) ◽  
pp. 235-241 ◽  
Author(s):  
Ryuji Hiramatsu ◽  
Sueharu Horinouchi ◽  
Teruhiko Beppu
Keyword(s):  
Secretory Pathway ◽  
Isolation And Characterization

scholarly journals The Blocked Amino-Terminal Peptide of Feather Keratin

1971 ◽  
Vol 24 (1) ◽  
pp. 179 ◽  
Author(s):  
IJ O'donnell
Keyword(s):  
Molecular Weight ◽  
Acetic Acid ◽  
Amino Groups ◽  
Amino Terminal ◽  
Acetyl Content ◽  
Electrophoretic Patterns ◽  
Isolation And Characterization ◽  
Primary Amino

Proteins extracted from reduced and carboxymethylated feather keratins (SCM-keratins) have been studied by Harrap and Woods (1964a, 1964b, 1967). They have demonstrated the presence of electrophoretic heterogeneity amongst the proteins and have obtained a molecular weight of approximately 11,000 in agreement with earlier work of Woodin (1954). There was no indication of marked heterogeneity with respect to size. Using acid hydrolysis and determination of acetic acid produced they found an acetyl content of 1 �30 molesj104 g in the rachis off owl feathers. These were thought to be attached to primary amino groups since there were no O-acetyl groups. In the present paper the isolation and characterization of the predominant, and probably sole, amino-terminal tripeptide from goose feather calamus is described. Goose feather calamus was chosen because its extracted proteins had one of the simplest electrophoretic patterns of proteins from the feathers of a number of species (Harrap and Woods 1967).

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