Documents about Budding Yeast

  • 8 Pages

    Lecture2_08_Handouts

    Virginia Tech, BIOL 2104

    Excerpt: ... Lecture 2 August 28, 2008 Multicellular Organisms; Model Organisms in Cell and Molecular Biology; Tissue Culture Cells Multicellular organisms are composed by multiple cells. Differentiated cells contain the information necessary for the development of a complete organism 1 Model Organisms Molecular biologists have focused on E. coli The budding yeast , Saccharomyces cerevisiae, is the model organism for eucaryotes 2 Arabidopsis thaliana is the model organism for plant The fruit fly Drosophila melanogaster occupies a central place in biological research The nematode worm Caenorhabditis elegans is one of the most used model systems in developmental biology 3 The frog Xenopus leavis is a commonly used species in biochemistry labs X. tropicalis X. leavis Mammals are much more complex than other animals Why? The mouse is the model organism for mammals 4 However, most of what we know about human cell biology has come from studies of human cells Fibroblasts Myoblasts Oligodendrocyte prec ...

  • 23 Pages

    LectureI

    Emory, IBS 501

    Excerpt: ... Cell Biology: Joshi Lecture I joshi@cellbio.emory.edu February 26, 2001 Teaching Assistant: Elaine Keane Cell Cycle I Reading Assignment: Chapter 17 in Alberts, 27 in Lewin, and 13 in Lodish Diverse experimental systems have been used to identify and isolate cell-cycle control proteins Xenopus laevis Cultured mammalian cells Clam eggs The budding yeast Saccharomyces cerevisiae The fission yeast Schizosaccharomyces pombe ...

  • 4 Pages

    04

    UC Davis, MIC 170

    Excerpt: ... MIC170 4/9/09 Lecture 4 (Thursday, 4/9) Yeast Cell - 5. Cytoskeleton, continued A. Actin B. Tubulin Heterodimer of and tubulins (~50 kDa each) polymerize to form microtubules Fig. 12.42 from Textbook -> yeast tubulins have 75% identical amino acid sequence to human tubulins "dynamic instability" in living cells, each microtubule elongates and shrinks very rapidly due to continuous polymerization/depolymerization (movie) Fig. 12.44 from the textbook Microtubule Function: i) Forms spindle that separate chromosomes in mitosis (appears to be the most important function) ii) Nuclear positioning at the bud neck before mitosis - Fig. 2 in handout-KS3 1 MIC170 4/9/09 Yeast Life Cycle We'll look at the life cycle of the budding yeast S. cerevisiae (Fig. 4.19, handout-4) Developmental options in budding yeast A. Vegetative growth Haploid (DNA content = 1n) or diploid (2n) B. Stationary phase C. Mating - two haploid cells of different mating types (a and ) conjugate and form diploid D. Meiosis and sp ...

  • 4 Pages

    15

    UC Davis, MIC 170

    Excerpt: ... ing -correlates with initiation of DNA replication (G1S) DNA staining by fluorescent dye, such as DAPI (4',6'-diamino-2phenylindole) nucleus at the bud neck - G2 to early M nuclear division - M Budding yeast has no distinct G2 phase - mitotic spindle appears before DNA replication is completed(S phase) B. measure DNA content DNA flowcytometry using fluorescence-activated cell sorter (FACS) Fig. 4-31 (handout-8) stain DNA of cells with fluorescent dye that specifically binds to DNA and measure the amount of fluorescence for each cell (~10,000 cells/experiment) Fig. 17-6 (handout-8) Also informative to know the length of each cell cycle phases of growing culture - length of each cell cycle phase can be calculated from the data 2 MIC170 5/21/09 Two essential tools for cell cycle study in yeast A. Synchronized culture B. cdc mutants A. Synchronized culture (to analyze events specific to each cell cycle phase) If you just inoculate some yeast cells in liquid medium and incubate, y ...

  • 6 Pages

    s2

    Berkeley, MCB 140

    Excerpt: ... NAME: _ STUDENT ID #: _ Very short answer questions (10 questions, 5 points each; 50 points total) Question 7. Leslie Orgel showed that a nonsense suppressor can be used to restore a modicum of inducibility to a particular lacI strain of E. coli. What was the (correct) interpretation of this result? Question 8. Even though the basic machinery of small-molecule-inducible gene regulation is very similar in E. coli and in budding yeast , the famous PaJaMo experiment would never work in yeast (or, for that matter, in any eukaryote) i.e., if done exactly as per Arthur Pardees outline. Why? Question 9. Yasuji Oshima incubated yeast cells carrying a gal4-ts allele in glucose at the restrictive temperature, then shifted the cells from the permissive to the restrictive temperature concomitant with the addition of galactose, and then showed that galactokinase synthesis was delayed relative to a wild-type control. What was the (correct) in ...

  • 5 Pages

    16

    UC Davis, MIC 170

    Excerpt: ... MIC170 5/26/09 Lecture 16 (Tuesday, 5/26) Continued on "Cell cycle control" Two essential tools for cell cycle study in yeast A. Synchronized culture B. cdc mutants B. cdc mutants cell division cycle mutants-arrest at a specific point in the cell cycle Many temperature-sensitive (ts) cdc mutants were isolated both in budding yeast (Lee Hartwell) and fission yeast (Paul Nurse). (a) Isolation of cdc mutants i. isolate conditional (ts) mutants - select for mutants that can grow at the permissive temperature (25oC) but not at the restrictive temperature (37oC) mutants of interest are those that show a specific block in cell cycle; all cells in the culture arrest at one certain point in the cell cycle with certain cytological landmark e.g., all cells arrest without bud - cdc mutant that arrests in G1 ii. cdc mutant is deficient in a gene product required to get the cell past the specific point in the cell cycle Fig. 17-25 (handout-8) example: a cdc mutant defective in the initiation of S phase at the r ...

  • 14 Pages

    Tanaka2005

    Princeton, MB 523

    Excerpt: ... REVIEWS KINETOCHORE CAPTURE AND BIORIENTATION ON THE MITOTIC SPINDLE Tomoyuki U. Tanaka, Michael J. R. Stark and Kozo Tanaka Abstract | Kinetochores are large protein complexes that are formed on chromosome regions known as centromeres. For high-fidelity chromosome segregation, kinetochores must be correctly captured on the mitotic spindle before anaphase onset. During prometaphase, kinetochores are initially captured by a single microtubule that extends from a spindle pole and are then transported poleward along the microtubule. Subsequently, microtubules that extend from the other spindle pole also interact with kinetochores and, eventually, each sister kinetochore attaches to microtubules that extend from opposite poles this is known as bi-orientation. Here we discuss the molecular mechanisms of these processes, by focusing on budding yeast and drawing comparisons with other organisms. ANEUPLOIDY A chromosome complement that is not a simple multiple of the haploid set. MITOTIC SPINDLE An intracel ...

  • 3 Pages

    07Lecture13

    Maryland, BSCI 410

    Excerpt: ... trein Summary of reporter genes Name Source Substrate 5-bromo-4-chloro 3-indoyl1-glucuronide (X-gluc) breakdown Visual Blue GUS (-glucuronidase) E.coli LacZ (-galactosidase) E. coli X-gal LUC (Luciferase) GFP Firefly luciferin & ATP none Blue emitting light Green (Green Flourescent Protein) Jelly fish High throughput analyses of the transcriptome Documenting gene expression on a genome wide scale Transcriptome: complete set of transcripts and their relative expression levels in a particular cell or tissue under defined conditions I. cDNA microarrays Use of DNA microarrays (chips) Fluorescently tagged cDNA probes are hybridized to DNA spots in the microarray for studying differential expression of thousands of genes at a time in two mRNA samples Fig. 12-21 II. Oligonucleotide microarrays (Affymetrix GeneChip) Light deprotection mask o o o o o OH OH o o o T T T o o o Sporulation gene expression profile in budding yeast Chu et al., (1998) Science 282, 699-705 Several classes of sporulation ...

  • 3 Pages

    CaseHistory6

    San Diego State, BIO 210

    Excerpt: ... Appendix N Case History # 6 Directions: Read the following case history and answer the questions, using your textbook, any available reference books and information from your lab exercises, as well as your notes from both the lab and lecture material and the Internet. (Questions = 0.5 point each, Total = 3 pts) A 25 year old woman who was being treated for breast cancer went to her doctor complaining of a sore mouth and difficulty swallowing. The doctor examined her mouth and found white patches on the inside as well as on her tongue. The white patches looked like cottage cheese or milk curds. The doctor was able to make his diagnosis based on the physical examination, but decided to take a culture and send it to the clinical laboratory for definitive identification. The laboratory made a wet mount of the sample and observed clusters of budding yeast cells, and pseudohyphae. Pseudohyphae are a series of buds remaining attached to the parent cell and appearing as filamentous hypha. The laboratory was able to m ...

  • 2 Pages

    lect4_questn wbg

    University of Florida, PCB 4522

    Excerpt: ... oteins or complexes must be present at the ARS in order for the Mcm2-7 complex to be loaded? 12. Which protein or complex utilizes ATP in loading the Mcm2-7 complex? 13. One (Cdc6 or Cdt1) is targeted for degradation in yeast and the other in metazoans. Which is which and what is the signal that initiates their being targeted? (Hint: see the slides entitled "Downregulation of licensing in late G1 phase.") 14. What comprises the licensing factor? 15. What is a good biological system to study licensing factor controls and why? 1 Version 4 16. Describe the role of proteins involved in the initiation of eukaryotic DNA replication. ORC proteins Cdc6, Cdt1 Mcm2-7 proteins 17. Which protein at the ARS has a short half life (~5 min) in budding yeast ? 18. Which protein complex at the ARS has helicase activity and unwinds the DNA ahead of the replication fork. (Note: It leaves the origin during S phase as it unwinds the DNA.) 19. Phosphorylation inhibits the loading activity of the ORC in all stages of the cell cyc ...

  • 6 Pages

    MCB 2007 November 14th Lecture Notes

    BC, BI 304

    Excerpt: ... s for cell-division cycle genes (cdc genes) o transformed yeast with every gene to identify which ones were responsible for cell cycle regulation o cdc2+ = wild type, cdc2- = wee phenotype (mutant, early entry into M phase) o code for the cdc2 protein o phenotype was easily observable in budding yeast s S. pombe and S. cerevisiae- cdc2 gene encodes for a CDK, cdc13 encodes a cyclin regulators: o cdc25 (decreases G2 length by stimulating transcription of cdc2) o wee1 (early entry into M via inhibition of cdc2 activity) regulatory genes are extremely conserved ...

  • 18 Pages

    EEB331-08_L12

    Toledo, EEB 331

    Excerpt: ... ci - yeast stage common; Recognized only recently from molecular evidence (Nishida and Sugiyama 1994): => their basal (ancient) nature is indicated from both molecular phylogenetic evidence and their simple life style and morphology. Saccharomycotina (= Hemiascomycetes) - lack ascomata; unitunicate asci - the "true" budding yeast s (-> Saccharomycetes, or the Saccharomycetales in traditional classification systems), but also includes filamentous forms in modern classification (-> Hemiasco-). Pezizomycotina (= Euascomycetes) - Ascomata observed when sexual stage known; - more complex morphology; -> `higher' ascomycetes - includes most lichen-forming fungi - includes ca. 90% of the known Ascomycota species Basidiomycota (a) Clark and Moncalvo, 2004 (b) Lutzoni et al., 2004 The Taphrinomycotina, if including Pneumocystis and Neolecta, is paraphyletic. Basidiomycota Note the discrepancy between mitochondrial and nuclear phylogenies at higher levels within the Ascomycota. - Is one tree wrong? - Are t ...

  • 13 Pages

    overview checkpoints

    UC Irvine, BC 207

    Excerpt: ... Review articles Checkpoints controlling mitosis Duncan J. Clarke1* and Juan F. Gimenez-Abian2 Summary Each year many reviews deal with checkpoint control.(1 5) Here we discuss checkpoint pathways that control mitosis. We address four checkpoint systems in depth: budding yeast DNA damage, the DNA replication checkpoint, the spindle assembly checkpoint and the mammalian G2 topoisomerase II-dependent checkpoint. A main focus of the review is the organization of these checkpoint pathways. Recent work has elucidated the order-of-function of several checkpoint components, and has revealed that the S phase, DNA damage and spindle assembly checkpoints each have at least two parallel branches. These steps forward have largely come from kinetic studies of checkpoint-defective mutants. BioEssays 22:351363, 2000. 2000 John Wiley & Sons, Inc. Introduction First, let us deal with the obligatory definition. A checkpoint is: `a mechanism that establishes a dependence relationship between two cellular processes ...

  • 2 Pages

    sp09_homework4

    Minnesota, CBS 4004

    Excerpt: ... Biol 4004-001, Spring 09 Homework 4 Name:_ 1. The budding yeast , Saccharomyces cerevisiae. has many advantages as an experimental system to study control of cell cycle. The cell cycle is completed in about 1.5 hours and the cell cycle stage can be determined from the shape of the cell, as shown in the drawing to the left. Assume that you have access to conditional cdc mutants in a haploid background. At the restrictive temperature, the protein encoded by the mutant gene loses its normal function. For the conditions listed below, draw a graph from flow cytometry analysis such as the one in textbook fig. 17-13 to show the expected DNA content of the cells. Please be sure that the graph accounts for all the cells in the population and is properly labeled. In a few words, please explain the results in your drawings. (6 points total) Lodish et al., Mol. Cell. Biol., fourth edition a) mutant cdc20 cells grown for 24 hr at the restrictive temperature b) 20 min after moving the cells to th ...

  • 3 Pages

    midtermkey2

    UC Davis, MIC 170

    Excerpt: ... MIC170 Midterm May 5, 2009 Part I. Choose the ONE most appropriate answer for each of questions 1~5. 1. (2 points) The lipid bilayer of the plasma membrane is NOT permeable to glucose 2. (2 points) You are planning to construct a genomic library of the budding yeast S. cerevisiae. If the average size of the genomic DNA fragment in each plasmid is about 10 kb, you would want to make a library containing at least ( ) different plasmids to make sure the complete coverage of the entire genome. 6,000 3. (2 points) You are about to start chemical mutagenesis of the budding yeast S. cerevisiae. As you learned in the class, it would be ideal that a single mutation causes an interesting phenotype. Theoretically, to induce one mutation per cell in S. cerevisiae, the mutation frequency by mutagenesis needs to be about; 6x10-3/gene 4. (3 points) It's almost 9PM and you want to go home. You have an exponentially growing yeast cell culture, whose current concentration is 1.6 x 106 cells/ml. It has been determined that yea ...

  • 10 Pages

    TanakaDiffley

    Princeton, MB 523

    Excerpt: ... articles Interdependent nuclear accumulation of budding yeast Cdt1 and Mcm27 during G1 phase Seiji Tanaka and John F.X. Diffley* Cancer Research UK, Clare Hall Laboratories, Blanche Lane, South Mimms, EN6 3LD, UK *e-mail: J.Diffley@cancer.org.uk Published online: 11 February 2002, DOI:10.1038/ncb757 Cdt1 is essential for loading Mcm27 proteins into prereplicative complexes (pre-RCs) during replication licensing and has been found in organisms as diverse as fission yeast and humans. We have identified a homologue of Cdt1 in Saccharomyces cerevisiae, which is required for pre-RC assembly. We show that, like Mcm27p, Cdt1p accumulates in the nucleus during G1 phase and is excluded from the nucleus later in the cell cycle by cyclin dependent kinases (cdks). Cdt1p interacts with the Mcm27p complex, and the nuclear accumulation of these proteins during G1 is interdependent. This coregulation of Cdt1p and Mcm27p represents a novel level of pre-RC control. T he assembly of pre-RCs at origins of DNA ...

  • 12 Pages

    Cromieetal2007

    UC Davis, MIC 275

    Excerpt: ... budding yeast Saccharomyces cerevisiae by electron microscopy (Bell and Byers, 1983) and deduced from two-dimensional (2D) gel analysis of DNA (Schwacha and Kleckner, 1995), and it has been widely assumed that double HJs are a universal precursor of crossovers. However, their existence has not, to our knowledge, been reported in any other organism. In meiotic recombination, joint molecules can form between sister chromatids or between homologous chromosomes since either can provide the sequence homology needed to repair a DSB. However, only interactions with the homolog can result in crossovers that reassort genetic information and aid correct segregation of chromosomes at MI. Consistent with the importance of interhomolog events, the results of a study in Locusta migratoria suggested that interhomolog crossovers outnumber sister chromatid exchanges as visualized by differential BrdU staining (Tease and Jones, 1979). More directly, in budding yeast it was observed that interhomolog joint molecules predominat ...

  • 7 Pages

    03

    UC Davis, MIC 170

    Excerpt: ... MIC170 4/7/09 Lecture 3 (Tuesday, 4/7) Yeast cell 1. 2. 3. 4. 5. Cell surface Nucleus Mitochondria Vesicular organelles Cytoskeleton 2. Nucleus (continued from the last lecture) Essential elements in yeast linear chromosomes 1) CEN 2) ARS 3) TEL 1) CEN (centromere) sequences - One per chromosome (handout KS-2) = Sites of kinetochore formation - microtubules attach to kinetochore to separate sister chromatids in mitosis Fig. 5.19 from Textbook -> In budding yeast , 125-bp DNA is sufficient for centromere function <- many different proteins bind to form kinetochore Comparison of centromere DNA structures (Fig. 5.21 from Textbook) the 1 MIC170 4/7/09 2) ARS (Autonomously Replicating Sequence) - ~100bp First identified as sequences that enable plasmid DNA to replicate autonomously (Fig. 6.14 from Textbook) turned out it contains replication origin ORC (origin recognition complex), which contains six different proteins, binds to origin to initiate DNA replication Budding yea ...

  • 3 Pages

    test1key

    UC Davis, MIC 170

    Excerpt: ... tion of - and -( tubulin ) heterodimer. This characteristic of microtubules is known as "( dynamic instability )". G. In budding yeast , birth of daughter cells leaves distinct structures on the mother cell surface. They are called ( bud scar(s) ). 2. (5 points) Write "True" or "False" beside each of the following statements about yeast nucleus. True False Nucleolus is the site of ribosomal RNA synthesis and ribosomal assembly. Mitosis in yeast and fungi is known as "closed mitosis" because their nuclear envelope breaks down during mitosis. In comparison with the budding yeast Saccharomyces cerevisiae genome, the human genome contains ~267 times more genes. Each yeast chromosome has one ARS, the site of kinetochore formation. Every end of yeast chromosomes has a TEL sequence. These TEL sequences are synthesized by a specialized RNA polymerase called "telomerase". False False False 3. The mating type of the budding yeast S. cerevisiae is determined by the MAT locus on chromosome III. (a) (4 points) As we'l ...

  • 1 Pages

    duncker

    W. Alabama, BIOL 499

    Excerpt: ... Biology 499 - Senior Honours Projects Dr. Bernard Duncker bduncker@sciborg.uwaterloo.ca Office: B1 293A Ext. 3957 My laboratory is using the budding yeast Saccharomyces cerevisiae to study protein factors which control the initiation of DNA replication. S. cerevisiae has proven to be a very useful organism for such studies because it is one of the few eukaryotes for which origins of replication have been well characterised, and the only one for which an origin consensus sequence has been identified. These findings, in combination with an advanced knowledge of budding yeast genetics, has permitted the identification of numerous protein factors which associate with replication origins. Studies with S. cerevisiae have revealed that a number of protein factors assemble at replication origins in G1-phase of the cell cycle, leading to the formation of the pre-replicative complex (pre-RC). The pre-RC must be present in order for origins to fire, and is rapidly disassembled in S-phase. Mechanisms have been discovere ...