CSB428H1 Study Guide - Immunoprecipitation, Epistasis, Cantharidin

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CSB428H1F Article #6: Krahn et al.Formation of Baz-Sdt for epithelial polarity
Introduction
Baz phosphorylation site bind PDZ of Sdt to recruit Sdt to PM.
Phosphorylation of Baz causes it to dissociate from Sdt. Non-phosphorylatable Baz blocks
dissociation causing mutation similar to
crb
and
sdt
.
Crb cytoplasmic domain binds PDZ domain of Sdt and Sdt recruit Patj and Lin-7 to apical complex.
Dlg/Lgl/Scrib localize to lateral PM and antagonize Crb-Sdt complex, restricting apical expansion.
BazS980 phosphorylation is required for localization
WT Baz-GFP and Baz
S980E
-GFP shows the same localization basal to Crb-Sdt.
Phosphorylated BazS980-P was concentrated in most apical part, sometimes with/without Crb-Sdt.
WT Baz-GFP and Baz
S980E
-GFP but not Baz
S980A
-GFP rescued maternal and zygotic
baz
lethality.
Overexpression of Baz
S980A
-GFP is similar to
crb and sdt
Overexpression of Baz
S980A
-GFP caused for mation of aggregate containing D E-Cad, Arm,
-cat,
Par-6, aPKC, Crb, Sdt, Patj and Lin7. Lateral marker Dlg was normal.
Baz
S980A
-GFP overexpression caused cells to round up and die of apoptosis.
These dominant-negative effects of Baz
S980A
-GFP were sell-autonomous. Deletion of Baz CR1 or
PDZ domains did not affect is dominancy. However, those lacking PM targeting signal was not
dominant, showing Baz
S980A
-GFP must be at PM to be dominant.
In neuroblasts and oocytes, Baz
S980A
-GFP localize apically like WT and cell fate determinants,
spindle orientation, cell division, and viability was not affected.
Baz
S980A
-GFP mimic crb/
sdt
crb
,
sdt
,
baz
,
aPKC
, and
Par-6
mutants secrete only little scraps of cuticle.
Overexpressing Baz
S980A
-GFP caused a cuticle phenotype similar to
crb
or
sdt
mutants, but
phenotype can be rescued with
lgl
,
dlg
and
scrib
mutation, showing Crb antagonizes Scrib complex.
Overexpressing intracellular por tion of Crb (Crb
intra
) caused ectopic cuticle release.
Overexpressing Baz
S980A
-GFP with Crb
intra
has the same phenotype as overexpressing Baz
S980A
-GFP
1
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Document Summary

baz phosphorylation site bind pdz of sdt to recruit sdt to pm. Phosphorylation of baz causes it to dissociate from sdt. Non-phosphorylatable baz blocks dissociation causing mutation similar to crb and sdt. crb cytoplasmic domain binds pdz domain of sdt and sdt recruit patj and lin-7 to apical complex. dlg/lgl/scrib localize to lateral pm and antagonize crb-sdt complex, restricting apical expansion. wt baz-gfp and bazs980e-gfp shows the same localization basal to crb-sdt. Phosphorylated bazs980-p was concentrated in most apical part, sometimes with/without crb-sdt. wt baz-gfp and bazs980e-gfp but not bazs980a-gfp rescued maternal and zygotic baz lethality. Overexpression of bazs980a-gfp is similar to crb and sdt. overexpression of bazs980a-gfp caused formation of aggregate containing de-cad, arm, -cat, bazs980a-gfp overexpression caused cells to round up and die of apoptosis. However, those lacking pm targeting signal was not dominant, showing bazs980a-gfp must be at pm to be dominant.

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