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CELL 301 > Part B: Lecture 9 + 10 + 11 > Flashcards

Part B: Lecture 9 + 10 + 11 Flashcards

1
Q

Ahmed paper summary

A

-identify two gene recruitment sequences (GRS I and II) from promoter of INO1 gene to target gene to nuclear periphery
-mechanism is highly conserved over evolution

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2
Q

Blobel proposed the ______ which stated that _____

A

-gene gating hypothesis
-actively transcribed genes are tethered to NPC

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3
Q

silenced heterochromatin is bound to the ________ but abruptly interrupted at the nuceloplasmic face of the NPC by the ______

A

-inner nuclear membrane
-presence of transcriptionally active euchromatin

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4
Q

Nups aka ____

A

nucleoporin

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5
Q

Nups aka ____

A

nucleoporin

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6
Q

yeast is a good model organism because _____

A

you can change growth media/environment/nutrient source to turn on/off genes

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7
Q

Turned on genes move to ____

A

NPC

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8
Q

Localization of a gene in nucleus can be examined in a living cell using a ____

A

tattooing procedure

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9
Q

chromosome tattooing summary

A

-GFP-tag on protein that binds to gene (ex. Lac repressor -> binds to Lac O array repeats)
-real-time fluorescence

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10
Q

Nup49-GFP illuminates the _____

A

nuclear periphery

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11
Q

INO1 gene function? turned on/off when?

A

-generation of inositol, important in. lipid genesis
-turned on in inositol-depleted media
-turned off in inositol-rich media

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12
Q

GRS I in promoter of INO1 gene is ____

A

8 bp long and targets to NPC

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13
Q

Interestingly, introduction of the grs I mutation into the promoter of the endogenous INO1 gene did not ______. This means that there is a ______

A
  • block targeting of the endogenous INO1 to the nuclear periphery
  • second targeting sequence upstream of the INO1 promoter, termed GRS II (not related to GRS I)
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14
Q

upstream of GRS I is the ____

A

GRS II

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15
Q

mutant GRS I & II = _____

A

no association with nuclear periphery

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16
Q

TSA2 gene encodes an ______ that is activated by ________

A

-inducible thioredoxin peroxidase
-heat shock and oxidative stress

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17
Q

_______ were dispensable for peripheral targeting of INO1. However, most of the proteins associated with the ______ were required for peripheral localization of INO1.

A

-Most proteins that make up the core channel of the NPC
-nucleoplasmic face of the NPC

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18
Q

GRS I from INO1 and TSA2 is related to a sequence motif that was previously found to be _____

A

overrepresented in promoters of NPC-associated genes

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19
Q

INO1 mRNA levels were quantified by _______ following induction by _______ from strains having either plasmid-borne _____ integrated at URA3.

A

-reverse transcription-qPCR
-inositol starvation
-wildtype (WT) INO1 or grs I mutant INO1

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20
Q

GFP-Sec61 is a marker for ______

A

ER/NE

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21
Q

changes to nuclear envelope during metaphase

A

post-translational modification drive disassembly done by phosphorylation

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22
Q

changes to nuclear envelope during anaphase/telophase

A

chromatin interactions with the nuclear envelope are re-established

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23
Q

phosphorylation of lamins and NPC proteins does what?

A

-disrupts a lot of protein-protein interactions
-forms nuclear envelope fragments during prophase

24
Q

dephosphorylation of lamins and NPC proteins does what?

A

-promote assembly of nuclear fragments into nucleus
-nuclear envelope fusion during telophase

25
Q

open mitosis def. + type of cells

A

-NE disrupted
-eukaryotic cells + metazoan cells

26
Q

closed mitosis def. + type of cells

A

-NE stays intact
-yeast cells

27
Q

_______ can be used to examine changes in nuclear envelope and chromatin interactions during mitosis because ______

A

-S. cerevisiae
-it uses closed mitosis; NE stays intact

28
Q

NE is continuous with _____

A

ER

29
Q

GFP Tel 14 is a marker of ____

A

NE

30
Q

GFP INO1 is a marker of ____

A

NPC

31
Q

_______ is a positive regulator of post mitotic nuclear envelope reassembly

A

SUMOylation

32
Q

SUMO aka _____

A

Small ubiquitin-like modification (15kDa); similar to ubiquitin but doesn’t cause degradation

33
Q

SUMO cycle

A

-SUMO E1 activating enzyme binds to SUMO then transfers to SUMO E2
-SUMO E2 conjugating enzyme; donor of SUMO to other targets
-SUMO E3 is a ligase that links SUMO from SUMO E2 to target
-target-SUMO has SUMO removed by SUMO proteases

34
Q

parts of SUMO E1

A

Uba2, Aos1

35
Q

parts of SUMO E2

A

Ubc9

36
Q

SUMO E3 ligase names

A

-Siz1, Siz2, Mms21

37
Q

SUMO proteases/isopeptidases names

A

Ulp1, Ulp2

38
Q

SUMOylaFon at the nuclear envelope ______ during mitosis.

A

increases

39
Q

The increase in SUMOylation at the nuclear envelope coincides with _______ to the nuclear envelope during mitosis.

A

Siz2 recruitment

40
Q

The mitotic enrichment of SUMOylation at the nuclear envelope is
dependent on _____._____ is enriched at the nuclear envelope during mitosis. The mitosis specific SUMOylation of several proteins is dependent on ___.

A

-Siz2

41
Q

Clb2 is a tracker of ____

A

mitosis

42
Q

Loss of Siz2 = ____

A

loss of SUMOlyation

43
Q

Phosphorylation of Siz2 at _______ is required for recruitment to the nuclear envelope during mitosis.

A

serine residue 522

44
Q

Scs2 is a _______

A

-a VAP (VAMP-associated protein) family member
-tail-anchored integral membrane protein present in the ER (SUMO site in cytoplasm) and the inner nuclear membrane (SUMO site in nucleoplasm)

45
Q

Scs2 is used as a ____

A

hook for other proteins

46
Q

General functions of Scs2

A

-ERAD, deubiquination
-lipid droplet
-plasma membrane tether of actin
-mitochondria tether

47
Q

Scs2 SUMO site is where?

A

-before the transmembrane region (C-terminal)
-SUMO site: VKKE (Val-Lys-Lys-Glu); SUMO is added here

48
Q

Scs2 N-terminus

A

MSP = + a.a residues; interacts with FFAT motif

49
Q

_____ is nuclear envelope receptor for Siz2 during mitosis.

A

Scs2

50
Q

Scs2, Siz2 interaction in late mitosis

A

-Siz2 is phosphorylated & SUMOlyates Scs2
-Siz2 FFAT motif interacts with Scs2 MSP motif (+++)
-SUMO site of Siz2 adds SUMO to Scs2

51
Q

end of mitosis: Scs2 remove ____; Siz2 _____

A

-SUMO
-dephosphorylated

52
Q

Recruitment of Siz2 to the nuclear envelope coincides with the reestablishment of _____

A

chromatin interactions with the nuclear envelope

53
Q

Siz2-mediated SUMOylation at the nuclear envelope facilitates the reassociation of ______

A

telomeres to the nuclear envelope during mitosis

54
Q

____ is required for the association of telomeres with the nuclear
envelope during mitosis

A

-Sir4

55
Q

Nuclear envelope recruitment of ____ and SUMOylation of ____ are required for telomere association with the nuclear envelope.

A

-Siz2
-Sir4

56
Q

chromatin IP (CHIP)

A

-pull down chromatin to see proteins associated with DNA

57
Q

In late mitosis, a cascade of _____ faciliates the re-assoaction of chromatin to nuclear envelope during mitosis + association of proteins associated with NE

A

SUMOlyation

CELL 301 (7 decks)

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