scholarly journals A novel factor essential for unconventional secretion of chitinase Cts1

2020 ◽  
Author(s):  
Michèle Reindl ◽  
Janpeter Stock ◽  
Kai P. Hussnaetter ◽  
Aycin Genc ◽  
Andreas Brachmann ◽  
...  
Keyword(s):  
Cell Division ◽  
Underlying Mechanism ◽  
Yeast Cells ◽  
Subcellular Targeting ◽  
Uv Mutagenesis ◽  
Daughter Cells ◽  
Secretion Pathway ◽  
Unconventional Secretion ◽  
Mother And Daughter ◽  
Two Hybrid

AbstractSubcellular targeting of proteins is essential to orchestrate cytokinesis in eukaryotic cells. During cell division of Ustilago maydis, for example, chitinases must be specifically targeted to the fragmentation zone at the site of cell division to degrade remnant chitin and thus separate mother and daughter cells. Chitinase Cts1 is exported to this location via an unconventional secretion pathway putatively operating in a lock-type manner. The underlying mechanism is largely unexplored. Here, we applied a forward genetic screen based on UV mutagenesis to identify components essential for Cts1 export. The screen revealed a novel factor termed Jps1 lacking known protein domains. Deletion of the corresponding gene confirmed its essential role for Cts1 secretion. Localization studies demonstrated that Jps1 colocalizes with Cts1 in the fragmentation zone of dividing yeast cells. While loss of Jps1 leads to exclusion of Cts1 from the fragmentation zone and strongly reduced unconventional secretion, deletion of the chitinase does not disturb Jps1 localization. Yeast-two hybrid experiments suggest that the two proteins interact. In essence, we identified a novel component of unconventional secretion that functions in the fragmentation zone to enable export of Cts1. We hypothesize that Jps1 acts as an anchoring factor, supporting the proposed novel lock-type mechanism of unconventional secretion.

scholarly journals A Mutation in the ATP2 Gene Abrogates the Age Asymmetry Between Mother and Daughter Cells of the Yeast Saccharomyces cerevisiae

Genetics ◽  
2002 ◽  
Vol 162 (1) ◽  
pp. 73-87 ◽  
Author(s):  
Chi-Yung Lai ◽  
Ewa Jaruga ◽  
Corina Borghouts ◽  
S Michal Jazwinski
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Division ◽  
Life Span ◽  
Mother Cell ◽  
Yeast Cells ◽  
Mitochondrial Morphology ◽  
Temperature Sensitive ◽  
Yeast Saccharomyces Cerevisiae ◽  
Daughter Cells ◽  
Mother And Daughter

Abstract The yeast Saccharomyces cerevisiae reproduces by asymmetric cell division, or budding. In each cell division, the daughter cell is usually smaller and younger than the mother cell, as defined by the number of divisions it can potentially complete before it dies. Although individual yeast cells have a limited life span, this age asymmetry between mother and daughter ensures that the yeast strain remains immortal. To understand the mechanisms underlying age asymmetry, we have isolated temperature-sensitive mutants that have limited growth capacity. One of these clonal-senescence mutants was in ATP2, the gene encoding the β-subunit of mitochondrial F1, F0-ATPase. A point mutation in this gene caused a valine-to-isoleucine substitution at the ninetieth amino acid of the mature polypeptide. This mutation did not affect the growth rate on a nonfermentable carbon source. Life-span determinations following temperature shift-down showed that the clonal-senescence phenotype results from a loss of age asymmetry at 36°, such that daughters are born old. It was characterized by a loss of mitochondrial membrane potential followed by the lack of proper segregation of active mitochondria to daughter cells. This was associated with a change in mitochondrial morphology and distribution in the mother cell and ultimately resulted in the generation of cells totally lacking mitochondria. The results indicate that segregation of active mitochondria to daughter cells is important for maintenance of age asymmetry and raise the possibility that mitochondrial dysfunction may be a normal cause of aging. The finding that dysfunctional mitochondria accumulated in yeasts as they aged and the propensity for old mother cells to produce daughters depleted of active mitochondria lend support to this notion. We propose, more generally, that age asymmetry depends on partition of active and undamaged cellular components to the progeny and that this “filter” breaks down with age.

scholarly journals Complex Minisatellite Rearrangements Generated in the Total or Partial Absence of Rad27/hFEN1 Activity Occur in a Single Generation and Are Rad51 and Rad52 Dependent

2006 ◽  
Vol 26 (17) ◽  
pp. 6675-6689 ◽  
Author(s):  
Judith Lopes ◽  
Cyril Ribeyre ◽  
Alain Nicolas
Keyword(s):  
Pedigree Analysis ◽  
Model System ◽  
Yeast Cells ◽  
Single Generation ◽  
Daughter Cells ◽  
Double Deletion ◽  
Mother And Daughter ◽  
Complex Events ◽  
High Degree ◽  
Insight Into

ABSTRACT Genomes contain tandem repeat blocks that are at risk of expansion or contraction. The mechanisms of destabilization of the human minisatellite CEB1 (arrays of 36- to 43-bp repeats) were investigated in a previously developed model system, in which CEB1-0.6 (14 repeats) and CEB1-1.8 (42 repeats) alleles were inserted into the genome of Saccharomyces cerevisiae. As in human cells, CEB1 is stable in mitotically growing yeast cells but is frequently rearranged in the absence of the Rad27/hFEN1 protein involved in Okazaki fragments maturation. To gain insight into this mode of destabilization, the CEB1-1.8 and CEB1-0.6 human alleles and 47 rearrangements derived from a CEB1-1.8 progenitor in rad27Δ cells were sequenced. A high degree of polymorphism of CEB1 internal repeats was observed, attesting to a large variety of homology-driven rearrangements. Simple deletion, double deletion, and highly complex events were observed. Pedigree analysis showed that all rearrangements, even the most complex, occurred in a single generation and were inherited equally by mother and daughter cells. Finally, the rearrangement frequency was found to increase with array size, and partial complementation of the rad27Δ mutation by hFEN1 demonstrated that the production of novel CEB1 alleles is Rad52 and Rad51 dependent. Instability can be explained by an accumulation of unresolved flap structures during replication, leading to the formation of recombinogenic lesions and faulty repair, best understood by homology-dependent synthesis-strand displacement and annealing.

scholarly journals A Potential Lock-Type Mechanism for Unconventional Secretion in Fungi

2019 ◽  
Vol 20 (3) ◽  
pp. 460 ◽  
Author(s):  
Michèle Reindl ◽  
Sebastian Hänsch ◽  
Stefanie Weidtkamp-Peters ◽  
Kerstin Schipper
Keyword(s):  
Cell Separation ◽  
Classical Pathway ◽  
Endomembrane System ◽  
Daughter Cells ◽  
Unconventional Secretion ◽  
Mother And Daughter ◽  
Complex Regulation ◽  
Higher Eukaryotes ◽  
Alternative Routes ◽  
Corn Smut

Protein export in eukaryotes can either occur via the classical pathway traversing the endomembrane system or exploit alternative routes summarized as unconventional secretion. Besides multiple examples in higher eukaryotes, unconventional secretion has also been described for fungal proteins with diverse functions in important processes such as development or virulence. Accumulating molecular insights into the different export pathways suggest that unconventional secretion in fungal microorganisms does not follow a common scheme but has evolved multiple times independently. In this study, we review the most prominent examples with a focus on the chitinase Cts1 from the corn smut Ustilago maydis. Cts1 participates in cell separation during budding growth. Recent evidence indicates that the enzyme might be actively translocated into the fragmentation zone connecting dividing mother and daughter cells, where it supports cell division by the degradation of remnant chitin. Importantly, a functional fragmentation zone is prerequisite for Cts1 release. We summarize in detail what is currently known about this potential lock-type mechanism of Cts1 secretion and its connection to the complex regulation of fragmentation zone assembly and cell separation.

scholarly journals Hsp104 Overexpression Cures Saccharomyces cerevisiae [ PSI + ] by Causing Dissolution of the Prion Seeds

2014 ◽  
Vol 13 (5) ◽  
pp. 635-647 ◽  
Author(s):  
Yang-Nim Park ◽  
Xiaohong Zhao ◽  
Yang-In Yim ◽  
Horia Todor ◽  
Robyn Ellerbrock ◽  
...  
Keyword(s):  
Molecular Chaperone ◽  
Fluorescent Protein ◽  
Yeast Cells ◽  
Yeast Prion ◽  
Yeast Prions ◽  
Daughter Cells ◽  
Amyloid Aggregates ◽  
Mother And Daughter ◽  
Green Fluorescent ◽  
Kinetics Of

ABSTRACT The [ PSI + ] yeast prion is formed when Sup35 misfolds into amyloid aggregates. [ PSI + ], like other yeast prions, is dependent on the molecular chaperone Hsp104, which severs the prion seeds so that they pass on as the yeast cells divide. Surprisingly, however, overexpression of Hsp104 also cures [ PSI + ]. Several models have been proposed to explain this effect: inhibition of severing, asymmetric segregation of the seeds between mother and daughter cells, and dissolution of the prion seeds. First, we found that neither the kinetics of curing nor the heterogeneity in the distribution of the green fluorescent protein (GFP)-labeled Sup35 foci in partially cured yeast cells is compatible with Hsp104 overexpression curing [ PSI + ] by inhibiting severing. Second, we ruled out the asymmetric segregation model by showing that the extent of curing was essentially the same in mother and daughter cells and that the fluorescent foci did not distribute asymmetrically, but rather, there was marked loss of foci in both mother and daughter cells. These results suggest that Hsp104 overexpression cures [ PSI + ] by dissolution of the prion seeds in a two-step process. First, trimming of the prion seeds by Hsp104 reduces their size, and second, their amyloid core is eliminated, most likely by proteolysis.

scholarly journals Spindle-independent condensation-mediated segregation of yeast ribosomal DNA in late anaphase

2005 ◽  
Vol 168 (2) ◽  
pp. 209-219 ◽  
Author(s):  
Félix Machín ◽  
Jordi Torres-Rosell ◽  
Adam Jarmuz ◽  
Luis Aragón
Keyword(s):  
Cell Division ◽  
Ribosomal Dna ◽  
Budding Yeast ◽  
Mitotic Cell ◽  
Yeast Genome ◽  
Daughter Cells ◽  
Late Anaphase ◽  
Mother And Daughter ◽  
The Right ◽  
Chromosome Resolution

Mitotic cell division involves the equal segregation of all chromosomes during anaphase. The presence of ribosomal DNA (rDNA) repeats on the right arm of chromosome XII makes it the longest in the budding yeast genome. Previously, we identified a stage during yeast anaphase when rDNA is stretched across the mother and daughter cells. Here, we show that resolution of sister rDNAs is achieved by unzipping of the locus from its centromere-proximal to centromere-distal regions. We then demonstrate that during this stretched stage sister rDNA arrays are neither compacted nor segregated despite being largely resolved from each other. Surprisingly, we find that rDNA segregation after this period no longer requires spindles but instead involves Cdc14-dependent rDNA axial compaction. These results demonstrate that chromosome resolution is not simply a consequence of compacting chromosome arms and that overall rDNA compaction is necessary to mediate the segregation of the long arm of chromosome XII.

scholarly journals Interaction of porcine circovirus-like virus P1 capsid protein with host proteins

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Libin Wen ◽  
Jiaping Zhu ◽  
Fengxi Zhang ◽  
Qi Xiao ◽  
Jianping Xie ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Underlying Mechanism ◽  
Yeast Cells ◽  
Cellular Proteins ◽  
Porcine Circovirus ◽  
Yeast Two Hybrid ◽  
Host Proteins ◽  
Host Interactions ◽  
Wasting Syndrome ◽  
Two Hybrid

Abstract Background Porcine circovirus-like virus P1 is a relatively new kind of virus that is closely related to the post-weaning multisystemic wasting syndrome, congenital tremors, and abortions in swine. The molecular mechanisms of P1 virus infection and pathogenesis are fully unknown. To analyze P1 and its host interactions, we used a yeast two-hybrid (Y2H) assay to identify cellular proteins interacting with the Cap of the P1 virus. In this study, the Cap of the P1 virus exhibited no self-activation and toxicity to yeast cells and was used as bait to screen the Y2H library prepared from the pancreas tissue. Results Five cellular proteins (EEP, Ral GDS, Bcl-2-L-12, CPS1, and one not identified) were found to interact with P1 Cap. The interaction between Cap and Ral GDS was confirmed by co-immunoprecipitation. Conclusions Our data are likely to support the future investigation of the underlying mechanism of P1 infection and pathogenesis.

scholarly journals Solid-phase inclusion as a mechanism for regulating unfolded proteins in the mitochondrial matrix

2020 ◽  
Vol 6 (32) ◽  
pp. eabc7288
Author(s):  
Linhao Ruan ◽  
Joshua T. McNamara ◽  
Xi Zhang ◽  
Alexander Chih-Chieh Chang ◽  
Jin Zhu ◽  
...  
Keyword(s):  
Solid Phase ◽  
Tricarboxylic Acid Cycle ◽  
Yeast Cells ◽  
Mitochondrial Proteins ◽  
Mitochondrial Matrix ◽  
Unfolded Proteins ◽  
Daughter Cells ◽  
Contact Sites ◽  
Age Dependent ◽  
Mother And Daughter

Proteostasis declines with age, characterized by the accumulation of unfolded or damaged proteins. Recent studies suggest that proteins constituting pathological inclusions in neurodegenerative diseases also enter and accumulate in mitochondria. How unfolded proteins are managed within mitochondria remains unclear. Here, we found that excessive unfolded proteins in the mitochondrial matrix of yeast cells are consolidated into solid-phase inclusions, which we term deposits of unfolded mitochondrial proteins (DUMP). Formation of DUMP occurs in mitochondria near endoplasmic reticulum–mitochondria contact sites and is regulated by mitochondrial proteins controlling the production of cytidine 5′-diphosphate–diacylglycerol. DUMP formation is age dependent but accelerated by exogenous unfolded proteins. Many enzymes of the tricarboxylic acid cycle were enriched in DUMP. During yeast cell division, DUMP formation is necessary for asymmetric inheritance of damaged mitochondrial proteins between mother and daughter cells. We provide evidence that DUMP-like structures may be induced by excessive unfolded proteins in human cells.

scholarly journals Mechanisms for Curing Yeast Prions

2020 ◽  
Vol 21 (18) ◽  
pp. 6536
Author(s):  
Lois E. Greene ◽  
Farrin Saba ◽  
Rebecca E. Silberman ◽  
Xiaohong Zhao
Keyword(s):  
Quality Control ◽  
Cell Division ◽  
Protein Quality ◽  
Prion Proteins ◽  
Amyloid Fibers ◽  
Yeast Prions ◽  
Daughter Cells ◽  
Mother And Daughter ◽  
Infectious Proteins ◽  
Β Sheet

Prions are infectious proteins that self-propagate by changing from their normal folded conformation to a misfolded conformation. The misfolded conformation, which is typically rich in β-sheet, serves as a template to convert the prion protein into its misfolded conformation. In yeast, the misfolded prion proteins are assembled into amyloid fibers or seeds, which are constantly severed and transmitted to daughter cells. To cure prions in yeast, it is necessary to eliminate all the prion seeds. Multiple mechanisms of curing have been found including inhibiting severing of the prion seeds, gradual dissolution of the prion seeds, asymmetric segregation of the prion seeds between mother and daughter cells during cell division, and degradation of the prion seeds. These mechanisms, achieved by using different protein quality control machinery, are not mutually exclusive; depending on conditions, multiple mechanisms may work simultaneously to achieve curing. This review discusses the various methods that have been used to differentiate between these mechanisms of curing.

scholarly journals Systematic analysis of asymmetric partitioning of yeast proteome between mother and daughter cells reveals “aging factors” and mechanism of lifespan asymmetry

2015 ◽  
Vol 112 (38) ◽  
pp. 11977-11982 ◽  
Author(s):  
Jing Yang ◽  
Mark A. McCormick ◽  
Jiashun Zheng ◽  
Zhengwei Xie ◽  
Mitsuhiro Tsuchiya ◽  
...  
Keyword(s):  
Cell Division ◽  
Asymmetric Cell Division ◽  
Asymmetric Distribution ◽  
Systematic Analysis ◽  
Daughter Cells ◽  
Yeast Proteome ◽  
Mother And Daughter ◽  
Asymmetric Partitioning ◽  
Mother Cells ◽  
Aging Factors

Budding yeast divides asymmetrically, giving rise to a mother cell that progressively ages and a daughter cell with full lifespan. It is generally assumed that mother cells retain damaged, lifespan limiting materials (“aging factors”) through asymmetric division. However, the identity of these aging factors and the mechanisms through which they limit lifespan remain poorly understood. Using a flow cytometry-based, high-throughput approach, we quantified the asymmetric partitioning of the yeast proteome between mother and daughter cells during cell division, discovering 74 mother-enriched and 60 daughter-enriched proteins. While daughter-enriched proteins are biased toward those needed for bud construction and genome maintenance, mother-enriched proteins are biased towards those localized in the plasma membrane and vacuole. Deletion of 23 of the 74 mother-enriched proteins leads to lifespan extension, a fraction that is about six times that of the genes picked randomly from the genome. Among these lifespan-extending genes, three are involved in endosomal sorting/endosome to vacuole transport, and three are nitrogen source transporters. Tracking the dynamic expression of specific mother-enriched proteins revealed that their concentration steadily increases in the mother cells as they age, but is kept relatively low in the daughter cells via asymmetric distribution. Our results suggest that some mother-enriched proteins may increase to a concentration that becomes deleterious and lifespan-limiting in aged cells, possibly by upsetting homeostasis or leading to aberrant signaling. Our study provides a comprehensive resource for analyzing asymmetric cell division and aging in yeast, which should also be valuable for understanding similar phenomena in other organisms.

Difference in generation times for mother and daughter cells in yeasts

1978 ◽  
Vol 24 (7) ◽  
pp. 827-833 ◽  
Author(s):  
T. W. Flegel
Keyword(s):  
Growth Characteristic ◽  
Time Lapse ◽  
Yeast Cells ◽  
Daughter Cells ◽  
Yeast Strains ◽  
Generation Times ◽  
Spot Check ◽  
Mother And Daughter ◽  
The Mean ◽  
Mother Cells

Fruitless attempts to synchronize haploid yeast cells from the Phragmobasidiomycete Sirobasidium magnum led to the discovery that the mother and daughter cells (MDC) had greatly different generation times. Time-lapse photographic sequences of budding showed that the mean generation time for daughter cells was more than three times greater than that for mother cells. This growth characteristic could be determined by a spot check of the microcolony pattern on agar. Using such a check, yeast strains of Rhodotorula (Rhodosporidium) and Cryptococcus that were tested demonstrated relative MDC equivalence while those of Sporobolomyces, Bullera, and Tremella showed MDC non-equivalence in varying degrees.

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