Beruflich Dokumente
Kultur Dokumente
By K. Sri Manjari
Research Scholar
Department of Cell Biology
Institute of Genetics & Hospital for Genetic Diseases
DBA is a
ribosomopathy
Mutations affecting
ribosomal protein
(RP) genes
DBA
Mutations affecting:
25-35%
25%
RPS19
40-50% ??
Heterozygous,
autosomal
dominant
Leading to RP
haploinsufficiency
Amino acids
RPS24
RPS7
RPS17
RPL26
RPS35a
RPL11
RPS10
Unknown
RPL5
RPS19
Standard
DBA
Screening
Pipeline
Measure & QC
Peripheral Blood
Extract DNA
RPS19
RPL5
RPL11
RPS24
RPS17
RPL35a
RPS7
Sanger Sequence
Darnell et al.,
2011
musashi-1 and -2
NPM1
hematological
malignancies
RPS7, RPS10,
RPS17, RPS19
DiamondBlackfan
anemia
TCOF1, POLR1D,
POLR1C
Treacher Collins
syndrome
DKC1
dyskeratosis congenita
THE PARADOX..
Individually, the ribosomopathies are rare and
phenotypically unique.
Intuitively, mutations affecting the ribosome, a
molecule essential for protein synthesis in every
cell, should affect all tissues and cell types.
On the contrary, ribosome biogenesis disorders
are highly heterogeneous in both their physical
manifestations and modes of inheritance, and
there is a surprising tendency toward tissue
specificity in these diseases
Over the last number of years, there has been increasing awareness of
the role that ribosome, ribosome biogenesis, and various other factors
that relate to translation play in normal cellular homeostasis, and in
human disease (Xue and Barna, 2012).
HYPOTHESIS
They hypothesize that the process of active
translation within the cell is regulated by a
multitude of proteins that can interact with
either the ribosome itself,
the mRNAs that are being actively translated,
or
proteins that may have the capacity to interact
with both the ribosome and mRNA.
EXPERIMENTAL PROCEDURES
EXPERIMENT
SILAC (stable isotope
labeling by amino acids in
cell culture) media to
incorporate amino acids for
light (Lys0 C13; Arg0 N14) or
heavy (Lys6 C13; Arg10 N15)
labeling of proteins,
achieving a labeling
efficiency of greater than
95%
Standard scatterplots with normalized Log2 (H/L) ratios/Log10 Intensities (Normal versus Cancer
n = 3, left panel; Cancer versus Cancer n = 3, middle panel;
PPC1 DMSO versus PP242, n = 3, right panel) highlighting the distribution of quantified proteins
in each screen (cutoff values for enriched proteins was 2 SDs (2s)
from the mean, dashed red lines). Proteins of interest in either experimental setting are
highlighted.
DISCUSSION
First, by cross-referencing data from independent SILAC
riboproteomic experiments showed that data set is highly
enriched in factors that relate directly to the ribosome, to
translational initiation and elongation, and to pathways that
are known to regulate and control translation.
Second, the data sets indicate that the diversity within the
riboproteome itself may have the capacity to categorize cell
types and tissues and, importantly, may specifically contribute
to regulation of gene expression within a given cellular
compartment.
DISCUSSION
Third, by examining globally how the riboproteome may be
altered in diseases such as human cancer, we have made
further unexpected observations. We find that riboproteomic
components display frequent copy-number amplifications in
human cancer, whereas genomic losses within the
riboproteome are significantly less than that for
nonriboproteomic genes.
Fourth, in addition to characterizing the riboproteome
landscape in various cell types, we identified and validated a
number of proteins previously not known to be associated
with actively translating ribosomes
Last, our data demonstrate that the cancer riboproteome can
be pharmacologically modulated for therapy on the basis of
this molecular knowledge.
Highlights
Mass spectrometry identifies differential
riboproteome components in cancer cells
Riboproteomic genes are frequent targets of
genetic amplification in cancer
Riboproteomics
identifies
previously
unrecognized ribosome-associated proteins
THANK YOU