Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic...
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Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression Fluorescences in the cytosol: Y-2A-C-2A-P DAPI 2A CMVp pA pac EYFP ECFP 2A T7p 2A pac EYFP ECFP 2A TRANSLATION mut FMDV 2A DAPI 2A CMVp pA pac EYFP ECFP 2A T7p 2A pac EYFP ECFP 2A TRANSLATION wt FMDV 2A
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Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression Problems in the ER translocon: G-Y-2A-C-2A-P 2A CMVp pA pacEYFPGTECFP 2A T7p TRANSLATION 2A pacEYFPGTECFP 2A DAPI ?
Transcript of Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic...
Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression
Fluorescences in the cytosol: Y-2A-C-2A-P
DAPI
2ACMVp pApacEYFP ECFP2AT7p
2A pacEYFP ECFP2A
TRANSLATION
mut FMDV 2A
DAPI
2ACMVp pApacEYFP ECFP2AT7p
2A pacEYFP ECFP2A
TRANSLATION
wt FMDV 2A
Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression
From the cytosol to the Golgi: Y-2A-G-C-2A-P
DAPI
2ACMVp pApacEYFP GT ECFP2AT7p
TRANSLATION
2A pacEYFP GT ECFP2A
Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression
Problems in the ER translocon: G-Y-2A-C-2A-P
2ACMVp pApacEYFPGT ECFP2AT7p
TRANSLATION
2A pacEYFPGT ECFP2ADAPI
?
Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression
More problems in the ER translocon: G-Y-2A-MT-C-2A-P
G-Y-2A-MT-C-2A-P 2ACMVp pApacEYFP MT ECFP2AT7p
TRANSLATION
2A pacEYFP MT ECFP2A
GT
GT
Y-2A-MT-C-2A-P 2ACMVp pApacEYFP MT ECFP2AT7p
TRANSLATION
2A pacEYFP MT ECFP2A
?
DAPI
Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression
Further problems in the translocon (I): G-Y-2A-G-C-2A-PG-Y-2A-G-C-2A-P
2ACMVp pApacEYFP GT ECFP2AT7p
TRANSLATION
2A pacEYFP GT ECFP2A
GT
GT
2ACMVp pApacECFP GT EYFP2AT7p
TRANSLATION
2A pacECFP GT EYFP2A
GT
GT
G-C-2A-G-Y-2A-P
? ?
Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression
Further problems in the translocon (II): Mitochondria!!
2ACMVp pApacECFP GT EYFP2AT7p
TRANSLATION
2A pacECFP GT EYFP2A
GT
GT
G-C-2A-G-Y-2A-PMT-C
CMVp pAMT ECFP
TRANSLATION
MT ECFP
Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression
A GalT problem in the polyprotein context?: ER-C-2A-G-Y
2ACMVp pAECFP GT EYFP
TRANSLATION
2AECFP GT EYFP
ER
ER
ER-C-2A-G-Y
2ACMVp pAECFP
TRANSLATION
2AECFP
ER
ER
ER-CN’
C
ER
CytosolN
N’
C
N
Type Itransmembrane secreted
Type II transmembrane
C
N
ER
Cytosol
?
