Proteinase function in yeast: Biochemical and genetic approaches to a central mechanism of post-translational control in the eukaryote cell

Rendueles, Paz Suárez; Wolf, Dieter H.

doi:10.1111/j.1574-6968.1988.tb02706.x-i1

Cited by 51 publications

(33 citation statements)

References 232 publications

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“…In eukaryotic cells two highly conserved degradation pathways exist: long-lived proteins are degraded within the lysosome, an organelle with membranes which protect the surrounding cytoplasm against lysosomal hydrolases ( Rendueles & Wolf, 1988); short-lived proteins are degraded by proteasomes, multimeric protease complexes which move between the nucleo- and cytoplasm ( Hershko & Ciechanover, 1998). Proteasomal substrates are often nuclear proteins such as proteins regulating cell cycle progression (cyclin-dependant kinases and their inhibitors), gene expression (transcriptions factors), DNA damage and stress response; although, misfolded proteins occurring during protein synthesis in the cytoplasm are also rapidly degraded by the proteasome ( Kirschner, 1999; Vabulas & Hartl, 2005; von Mikecz, 2006).…”

Section: Introductionmentioning

confidence: 99%

Intracellular Dynamics of the Ubiquitin-Proteasome-System

Chowdhury

Enenkel

2015

F1000Res

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The ubiquitin-proteasome system is the major degradation pathway for short-lived proteins in eukaryotic cells. Targets of the ubiquitin-proteasome-system are proteins regulating a broad range of cellular processes including cell cycle progression, gene expression, the quality control of proteostasis and the response to geno- and proteotoxic stress. Prior to degradation, the proteasomal substrate is marked with a poly-ubiquitin chain. The key protease of the ubiquitin system is the proteasome. In dividing cells, proteasomes exist as holo-enzymes composed of regulatory and core particles. The regulatory complex confers ubiquitin-recognition and ATP dependence on proteasomal protein degradation. The catalytic sites are located in the proteasome core particle. Proteasome holo-enzymes are predominantly nuclear suggesting a major requirement for proteasomal proteolysis in the nucleus. In cell cycle arrested mammalian or quiescent yeast cells, proteasomes deplete from the nucleus and accumulate in granules at the nuclear envelope (NE) / endoplasmic reticulum (ER) membranes. In prolonged quiescence, proteasome granules drop off the NE / ER membranes and migrate as stable organelles throughout the cytoplasm, as thoroughly investigated in yeast. When quiescence yeast cells are allowed to resume growth, proteasome granules clear and proteasomes are rapidly imported into the nucleus.Here, we summarize our knowledge about the enigmatic structure of proteasome storage granules and the trafficking of proteasomes and their substrates between the cyto- and nucleoplasm.Most of our current knowledge is based on studies in yeast. Their translation to mammalian cells promises to provide keen insight into protein degradation in non-dividing cells which comprise the majority of our body’s cells.

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Section: Introductionmentioning

confidence: 99%

Intracellular Dynamics of the Ubiquitin-Proteasome-System

Chowdhury

Enenkel

2015

F1000Res

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“…SDS. A. niger strain N647 was transformed as described by Tilburn et al (1983), using 5 pg (linearized) DNA.…”

Section: Molecular Methodsmentioning

confidence: 99%

“…6 7 2 3 49*4 1 5 t 2 1 7 2 4 9 6 2 4 6 1 5 6 1 8 2 2 101 5 s 12t-4 91 2 4 23 f 3 2 8 2 5 9 8 2 2 101 2 4 3 0 2 5 1 0 6 2 3 ~~ 4 9 2 4 103 -C 3 8 2 2 3 8 8 2 3 9 6 2 5 9 7 2 4 9 2 2 3 9 5 2 2 91 2 4 100 * s 102 2 2 97 f 2 103 2 4 106+3 100 + 6 1 0 2 5 4 differences have also been observed. In S. cerevisiae, it has been shown by Suarez Rendueles and Wolf (1988) that several vacuolar proteases are needed for the differentiation process of sporulation. They most likely provide the amino acids at the expense of unneeded (vegetative) cell protein for new protein synthesis for the endospores to be formed.…”

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confidence: 99%

Disruption of Three Acid Proteases in Aspergillus Niger— Effects on Protease Spectrum, Intracellular Proteolysis, and Degradation of Target Proteins

Hombergh¹,

Gelpke²,

Vondervoort³

et al. 1997

European Journal of Biochemistry

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Three acid protease genes encoding two extracellular proteases (PEPA and PEPB) and one intracellular protease (PEPE) were disrupted in Aspergillus niger. Northern-blot analysis showed the absence of wild-type protease mRNAs in the disruptants while western-blot analysis proved the absence of the encoded proteases. Characterization of the residual proteolytic spectra in the disruptants indicated that the extracellular protease activity was reduced to 16% and 94% for the ApepA and the ApepB disruptants, repectively. In the ApepE disruptant, the total intracellular proteolytic activity was reduced to 32 %. Apart from the reduced intracellular pepstatin-inhibitable aspartyl protease activity, serine protease and serine carboxypeptidase activities were also significantly reduced in the ApepE strain. This may indicate the presence of a cascade activation mechanism for several vacuolar proteases, triggered by the PEPE protein, similar to the situation in Saccharomyces cerevisiae. Disruption of a single protease gene had no effects on the transcription of other non-disrupted protease genes in A. niger. In supernatants of the disruptants, reduced degradation of a proteolytically very susceptible tester protein (PELB) was observed. By recombination, we also constructed ApepAApepB, ApepBApepE and ApepAApepE double disruptants as well as a ApepAApepBApepE triple disruptant, lacking all three acid protease activities. The in vitro residual PELB activity was the highest in the triple disruptant and the ApepAApepB recombinant.Keywords: Aspergillus ; protease ; disruption ; cascade activation ; proteolytic degradation.Aspergillus niger produces a broad spectrum of proteases, both specific and non-specific, which is recognized to be one of the major reasons for the low protein expression yields that are frequently observed, especially when producing heterologous proteins. The genetics of proteolytic degradation in Aspergilli is very complex due the involvement of many regulatory factors in protease expression. Cohen (1972Cohen ( , 1973 showed in a series of pioneering studies that in A. niduluns extracellular proteases were repressed by low-molecular-mass sources of carbon, nitrogen, and sulphur. and van den Hombergh et al. (1994) have recently shown that in A. niger both pathway-specific induction by proteins and wide domain regulatory mechanisms (carbon catabolite repression, nitrogen metabolite repression, and pH regulation) are involved in the overall regulation of extracellular proteases. Several intracellular

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“…Several intracellular proteolytic enzymes have been found in Saccharomyces cerevisiae 95,98 : two endoproteinases, proteinase A 2,20,21,62,69 and B 20,34,58 , two carboxypeptidases, carboxypeptidase Y and carboxypeptidase S 107 , several aminopeptidases 66,67 , and one dipeptidase 25 . The two main vacuolar endopeptidases yscA and yscB are essential for protein degradation during vegetative growth and under starvation con-ditions 74,80,101,106,108 .…”

Section: Intracellular Proteinase Activitymentioning

confidence: 99%

The Potential of Confocal Imaging for Measuring Physiological Changes in Brewer's Yeast

Schlee

Miedl

Leiper

et al. 2006

Journal of the Institute of Brewing

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This study focused on the implementation of fluorescence optical methods and laser scanning confocal microscopy for monitoring brewing yeast performance. Physiological parameters and cell compounds in yeast cells (glycogen, neutral lipids, trehalose, bud scars, DNA and intracellular proteinases) have been successfully visualised with the aid of highly specific fluorochromes. The expression and sub cellular localisation of proteinase A during fermentation has been studied employing a Saccharomyces cerevisiae green fluorescent protein clone. This novel approach to monitoring brewing yeast performance provides new insights into physiological events that occur during wort fermentation.

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Proteinase function in yeast: Biochemical and genetic approaches to a central mechanism of post-translational control in the eukaryote cell

Cited by 51 publications

References 232 publications

Intracellular Dynamics of the Ubiquitin-Proteasome-System

Intracellular Dynamics of the Ubiquitin-Proteasome-System

Disruption of Three Acid Proteases in Aspergillus Niger— Effects on Protease Spectrum, Intracellular Proteolysis, and Degradation of Target Proteins

The Potential of Confocal Imaging for Measuring Physiological Changes in Brewer's Yeast

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