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BMS242 CELL AND MOLECULAR > YEAST SCREENS > Flashcards

YEAST SCREENS Flashcards

1
Q

EUKARYOTIC CELLS NEED MEMBRANE TRAFFICKING BECAUSE A FEATURE OF EUKARYOTES IS …………………………

A

COMPARTMENTALISATION

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2
Q

COMPARTMENTALISATION ALLOWS MORE ………………………….

A

COMPLEXITY

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3
Q

CERTAIN ENZYMES REQUIRE CERTAIN …………………….. TO FUNCTION IN SUCH AS …………

A

ENVIRONMENTS

PH

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4
Q

WHAT ARE THE MAJOR FEATURES OF TRAFFICKING PATHWAYS

A
  1. PROTEIN SYNTHESIS ON BOUND RIBOSOMES - COTRANSLATIONAL TRANSPORT OF PROTEINS INTO/ACROSS ER MEMBRANE
  2. BUDDING/FUSION OF ER - GOLGI VESICLES FORM CIS GOLGI
  3. CISTERNAL PROGRESSION OR RETROGRADE GOLGI TO ER TRANSPORT
  4. RETROGRADE FROM LATER TO EARLIER GOLGI CISTERNAE
  5. CONSTITUITIVE SECRETION
  6. REGULATED SECRETION
  7. SORTING TO LYSOSOMES
  8. ENDOCYTOSIS
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5
Q

MODEL SYSTEMS FOR MEMBRANE TRAFFICKING REQUIRE WHAT

A

SIMPLE CELLULAR SYSTEM

A SYSTEM THAT HAS THE FUNCTION BEING STUDIED.

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6
Q

WHAT ARE THE ADVANTAGES OF YEAST AS A MODEL

A 
ENTIRE GENOME KNOWN
CAN GROW AS HAPLOID/DIPLOID
CHEAP AND EASY TO GROW IN LARGE AMOUNTS
LIMITED GENE DIVERSITY
FUNDAMENTAL PATHWAYS CONSERVED
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7
Q

WHAT ARE THE DISADVANTAGES OF YEAST AS A MODEL

A

LIMITED CELL-CELL CONTACT THEREFORE CANNOT STUDY MULTICELLULARITY
SMALL SO MICROSCOPY DIFFICULT
CELL WALL WHICH CAN MAKE SOME STUDIES DIFFICULT

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8
Q

WHO IDENTIFIED THE GENES OF THE SECRETORY PATHWAY

A

NOVICK AND SCHEKMAN 1980

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9
Q

WHAT WAS THE RATIONALE BEHIND THE INDENTIFICATION OF GENES IN THE SECRETORY PATHWAY

A

IF CELLS ARE SECRETORY DEFICIENT THEN THE CELL WOULD INCREASE IN DENSITY AS THE VESICLES CARRYING THE PROTEINS ACCUMULATE

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10
Q

WHAT ARE DEFINED SECRETORY MUTANTS

A

THOSE THAT FAIL TO EXPORT ACTIVE INVERTASE AND ACID PHOSPHATASE BUT CONTINUE TO SUNTHESISE PROTEIN

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11
Q

HOW MANY GENES WERE IDENTIFIED BY GROUPING MUTANTS BY PHENOTYPE IN MUTANT SECRETORY PROTEINS

A

23

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12
Q

ALPHA FACTOR IS GLYCOSYLATED AND …………………………….. CLEAVED AT DIFFERENT STAGES IN THE SECRETORY PATHWAY. BY FOLLOWING ITS PROCESS WE CAN LOCATE THE POSITION OF THE ……………………
THE SIZE OF THE PROTEIN (SHOW BY ………………………. BLOT) CAN SHOW WHICH STAGE OF ………………………….. ALPHA FACTOR GO TO.

A

PROTEOLYTICALLY
MUTATION
WESTERN
MODIFICATION

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13
Q

WHERE IS THE DECISION MADE AS TO WHETHER SOMETHING IS TRAFFICKED TO THE MEMBRANE OR TO A LYSOSOME

A

THE TRANS GOLGI NETWORK

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14
Q

HOW MANY CLASSES DO THE 23 MUTANT GENES IN THE SECRETORY PART OF THE PATHWAY FALL INTO

A

5

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15
Q

WHAT ARE THE CLASSES OF SEC MUTANTS

A

CLASS A: ACCUMULATION IN CYTOSOL
CLASS B: ACCUMULATION IN RER
CLASS C: ACCUMULATION IN ER TO GOLGI VESICLES
CLASS D: ACCUMULATION IN GOLGI
CLASS E: ACCUMULATION IN SECRETORY VESICLES

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16
Q

IN THE NOVICK SCHEKMAN EXPERIEMENTS WHAT ARE THE REASONS NOT EVERYTHING WAS IDENTIFIED

A

ONLY IDENTIFIED TEMPERATURE SENSITIVE MUTANTS
ONLY CONSIDERED THE SECRETORY PATHWAYS TO THE MEMBRANE SO DEFECTS IN ENDOSOME/VACUOLE WOULD NOT BE IDENTIFIED
REDUNDANT FUNCTIONING GENES (OVERLAP) WOULD NOT BE IDENTIFIED

17
Q

WHAT ARE THE TYPES OF SCREENING THAT CAN BE DONE IN MEMBRANE TRAFFICKING

A

SEC SCREENS
END SCREENS
VPS SCREENS

18
Q

WHAT IS ENDOCYTOSIS

A

PLASMA MEMBRANES INVAGINATE RESULTING IN A VESICLE THAT CAN FUSE WITH ENDOSOMES

19
Q

WHAT IS THE IMPORTANCE OF ENDOCYTOSIS

A

RETRIEVAL OF MOLECULES FOR RECYCLING
DOWN REGULATION OF SIGNALS
REMODELLING OF CELL SURFACE COMPOSITION
MEANS OF ENTRY BY TOXINS/PATHOGENS

20
Q

WHAT IS THE NAME OF ENDOCYTOTIC MUTANTS IN THE TRAFFICKING PATHWAY

A

END-

21
Q

HOW DO YOU DETECT END - MUTANTS

A

CANNOT INTERNALISE A FLUID PHASE MARKER SUCH AS LUCIFER YELLOW
IN WILD TYPE, WOULD LIGHT UP IN THE MIDDLE

22
Q

HOW MANY GENES HAVE BEEN DETECTED AS END - MUTANTS

A

7

23
Q

OF THE SEVEN END - MUTANTS, HOW MANY ARE DIRECTLY INVOLVED IN MEMBRANE INVAGINATION AND SCISSION

A

5

24
Q

WHAT IS THE FUNCTION OF A LYSOSOME

A

DEGRADES EC MATERIAL TAKEN UP BY ENDOCYTOSIS AS WELL AS SOME IC COMPONENTS

25
Q

WHY DO LYSOSOMES EXIST

A

ENZYMES MUST BE CONTAINED WITHIN THEIR OWN COMPARTMENT

26
Q

CARBOXYPEPTIDASE Y IS NORMALLY TRANSPORTED FROM ……………….. TO LYSOSOME. GENERATION OF MUTAGENISED CELLS AND MICROSCOPY IDENTIFIED OVER ………….. VPS GENES. MANY OF THESE GENES WORK IN ……………………
LIKE ALPHA FACTOR, CPY IS …………………… AND CLEAVED AND THIS CAN HELP IDENTIFY WHERE THE DEFECT HAS OCCURED.

A

GOLGI
60
COMPLEXES
GLYCOSYLATED

27
Q

VACUOLAR MUTATANT ARE DIVIDED INTO CLASSES DEPENDING ON THE STAGE OF BLOCKAGE. WHAT ARE THESE

A
  1. EARLY GOLGI
  2. LATE GOLGI
  3. EARLY ENDOSOME
  4. LATE ENDOSOME
  5. MVB
  6. LYSOSOME
28
Q

AT WHICH STAGE DO VACUOLAR MUTANTS OCCUR MOST

A

BETWEEN LATE ENDOSOME AND MVB

29
Q

MOLECULAR DETERMINANTS TELL US WHERE THE PROTEIN NEEDS TO GO. FROM THE TGN THERE ARE FOUR POSSIBLE DESTINATIONS. WHAT ARE THEY

A

PLASMA MEMBRANE
EARLY ENDOSOME
LATE ENDOSOME/MVB
VACUOLE

30
Q

HOW DO PROTEINS GET TO WHERE THEY NEED TO GO

A

IF THERE IS NO OR A DISRUPTED SIGNAL THEN CONTENTS WILL BE TRAFFICKED TO THE MEMBRANE

31
Q

WHAT IS THE CPY TRAFFICKING PATHWAY

A

CPY RECOGNISED BY RECEPTOR VPS10
THIS COMPLEX RECOGNISED BY CLATHRIN, GGA1/2
THIS SIGNALS THAT THIS WILL BE GOING TO THE LATE ENDOSOME
VPS10 DISSOCIATES FROM CPY AT THE LATE ENDOSOME
VPS10 IS RETRIEVED TO THE LATE GOLGI BY YSSL
CPY IS TRANSPORTED TO VACUOLES.

BMS242 CELL AND MOLECULAR (14 decks)

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