nukleoskelet a chromosomální domény

NUKLEOSKELET A
CHROMOSOMÁLNÍ DOMÉNY
Metodické těžkosti
HeLa.Fix, Extract, Fix, Fracture,FESEM
Nukleoskelet
Morfologie
In Vitro Nucleus, cryo-planing, FESEM
In Vitro Nucleus, cryo-abraded,FESEM
A living epidermal cell (PAM)
expressing GFP-lamin
before bleach
60 min after bleach
Moir et al. 2000
In Vitro Nucleus, DNA’ase, cryo-abraded,FESEM
In Vitro Nucleus, DNA’ase, cryo-abraded,FESEM
Mitotic chromosomes in scanning electron
microscope
Levels of
chromatin
condensation
Nukleoskelet
Složení
Nukleoskelet
Složení
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•
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lamins
hnRNPs
actin
NuMa
Nukleoskelet
Jaderný aktin a myosin
Actin shuttles through the nucleus
control
NES-mutated actin
Mapping the major difference peak
between ADP actin and NFA filaments
Buněčné jádro obsahuje myosin I
NMI obsahuje unikátní prodloužení o 16 aminokyselin na N-konci
Některé funkce myosinu I a myosinu II
v eukaryontních buňkách
Kolokalizace RNA pol II a NMI v
nukleoplasmě HeLa buněk
kontrola
α-amanitin
AMD
P<0.01
NS
NS
Anti-NMI protilátky inhibují transkripci
pomocí RNA pol II in vitro
Muscle contraction vs. transcription
Jaderný myosin-GFP
v živých 3T3 buňkách
Funkce jaderného skeletu
TRANSCRIPTION
REPLICATION
RNA pol II transcription
Lamin speckles form enlarged foci
upon "-amanitin treatment.
The Journal of Cell Biology, Volume 159, Number 5, 2002
Localization of lamin speckles and SFCs in
enlarged foci upon transcription inhibition
The Journal of Cell Biology, Volume 159, Number 5, 2002
Chromosomální a
interchromosomální domény.
Chromosome territories
chromosome 1
X chromosome
chromosome 17
Interchromatin
compartment
HeLa
GFP-H2B
SC-35
Expansion of less
condensed chromatin
into ICHC
Chromatin structure in situ (ESI)
Proteins chromatin+RNA
Nitrogen
Phosphorus
Gene-rich (chr. 19) and gene-poor
(chr. 18) chromosome territories
Xa and Xi territories from a
human female fibroblast nucleus
ANT2 and ANT3 (adenosine nucleotide translocase) genes
The multiloop
subcompartment model
Two 1-Mb chromatin domains or 'subcompartments' are shown
linked by a chromatin fibre. Each 1-Mb chromatin domain is
built up as a rosette of looped 100-kb chromatin fibres. Loops
are held together by a LOOP BASE SPRING, which simulates the
function of CHROMOSOME TERRITORY ANCHOR PROTEINS
Patterns of chromosome
positioning in mammalian cells
anaphase
telophase
Functional role for
chromosome positioning
END