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central ring
holds NPC in the nuclear envelope; transmembrane proteins
glycosylation
sugar chains added post-transcriptionally to NPC nucleoporins to face cytosol; block entry by adding drugs to mess up glycosylated residues
lectins
plant proteins with high affinity for sugar chains; ex. WGA; if injected into cells lectins bind to the nuclear pore and block entry/exit; used to study nuclear/cytoplasm exchange
nucleoplasm
large 145,000 kDA protein; head/tail domains, used to study NES/NLS
NLS/NES
basic (positively charged: lots of lysine) AA residues that are necessary and sufficient for cellular localization
digitonin
detergent, causes holes in plasma membrane and makes it completely permeable=all the cytosol leaks out
Rcc1
"regulator of chromosome condensation"; RanGEF, found exclusively in the nucleus of a cell
RanBP2
found at the entrance of the cytoplasm, nucleoporin, binds RanGTP
NTF2
nuclear transport factor, shuttles RanGDP into the nucleus, fastest shuttling protein in the cells
importin alpha
adaptor protein, recognizes NLS on cargo
importin beta
imports cargo into the nucleus
CAS
exports importin alphas into cytoplasm
CRM1
main exporter of most proteins and many ribonuclear complexes; if it is killed the cell will die
LMB
cytotoxic drug, blocks the nuclear export of all proteins (HIV Rev), binds to pocket of CRM1 that recognizes NES
HIV Rev
exports unspliced HIV RNA into the cytoplasm for virion packaging, contains RNA binding domain which is also an NLS to recog. a short seq. in RNA, and an NES, lots or arginine (positively charged) to bind to DNA; NOT transporter
heterokaryon
fused cells with shared cytoplasm and separate nuclei, can be 2 different species
core promoter
regulates transcription of gene by recruiting factors that initiate transcription, common to all genes that are being transcribed by RNA pol II, not sufficient to sustain transcription because of chromosome architecture
regulatory promoter
where TF associate to assist in transcription
enhancer
increase rate of transcription by making association of RNA pol II and core promoter as stable as possible, can be anywhere
Mediator
protein that helps TF associate with RNA pol II
pre-initiation complex
RNA pol II, TFs, and Mediator
TF
recruit holoenzyme to DNA, has DNA binding domain to bind to specific sites, protein binding domain for interaction with pre-initiation complex to activate transcription
H1
histone that sits at entry of DNA that wraps around histone, critical for 30 nm packing
histones
H2A&H2B dimers on the outside, more mobile; H3&H4 dimers on the inside; alpha helices interact with minor groove of DNA via arginine residues which are basic as opposed to DNAs acidity
acetylation
addition of acetyl group to lysine residues of histone tails in order to neutralize their positive charge and make them loosen their hold on the DNA; induce formation of euchromatin and therefore transcription; mostly H4
lamin
give structure to nucleus and organize heterochromatin; intermediate filaments, only one to have NLS; have long alpha helical structure that allows them to homo/heterodimer through coiled coil; leucine rich; not polar; not static
lamin B
present in embryos, if there is a mutation in them the cell will die, remain attached to cell membrane because their farnesyl group is not removed (has CaaX motif); 2 genes
lamin A
can have mutations; released as dimers; one gene--if cleaved with CaaX motif it is lamin A, if without it is lamin C which can only go through the membrane when attached to lamin A; can be in nucleoplasm=more roles than B; have farnesyl group cleave
LBR
lamin B receptor, interacts with heterochromatin via nucleoplasmic domain
LAP1
interacts with lamin B and A
LAP2
interacts with lamin A
HO gene
endonuclease; SWI2 mutant; mutant is suppressed by mutations in H4=weakens interactions between H4 and DNA
SW12/SNF
part of complex that disrupts/reconfigures nucleosomes and stabilizes the binding of TFs to nucleosomal DNA; if it is mutated the complex loses function; contains an ATPase; UNWINDS DNA from nucleosome (not slide)
SWI5
TF that recognize binding site in heterochromatin and recruits SWI2 complex; exclusive to HO gene; has DNA binding and protein binding domains
SAGA
histone acetylase, recruited by SWI/SNF complex
SBF
transcription factor recruited by SAGA that recruits general transcription factors
HATs
add acetyl groups
HDACs
remove acetyl groups
Gcn5
part of HAT complex, in yeast, transcriptional co-activator (you need it for transcription but its not sufficient alone)
Gcn4
TF, binds to UAS of DNA, recruits HAT complex, has DNA binding domain and protein binding domain
Ume6
recognizes URS of DNA, recruits Rpd3 HDAC via interaction with Sin3
5' Cap
occurs shortly after translation, 5' end recognized by cap binding complex which adds a methyl guanine cap
RNP
ribonucleoprotein, mRNA with associated proteins, spliced, capped, poly-A tail with poly-A binding proteins, ready for export
CTE
constitutive transport element, part of RNA, found in lower viruses, needs protein to carry it out of cell
Tap
heterodimer with P15, exports mRNA, help viral and cellular RNAs, coded for in cells, interact with nucleoporins, brought to mRNA via other RNAs that are in association
EJCs
exon junction complexes, deposited between exons by splicesome, recruit Tap and P15
eIF4E
eukaryotic initiation factor, replaces 5' cap, associates in cytoplasm after diffusion

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