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
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
?
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