843 research outputs found
MicroRNA-125b transforms myeloid cell lines by repressing multiple mRNA
Background: We previously described a t(2;11)(p21;q23) chromosomal translocation found in patients with myelodysplasia or acute myeloid leukemia that leads to over-expression of the microRNA miR-125b, and we showed that transplantation of mice with murine stem/progenitor cells overexpressing miR-125b is able to induce leukemia. In this study, we investigated the mechanism of myeloid transformation by miR-125b.
Design and Methods: To investigate the consequences of miR-125b over-expression on myeloid differentiation, apoptosis and proliferation, we used the NB4 and HL60 human promyelocytic cell lines and the 32Dclone3 murine promyelocytic cell line. To test whether miR-125b is able to transform myeloid cells, we used the non-tumorigenic and interleukin-3-dependent 32Dclone3 cell line over-expressing miR-125b, in xenograft experiments in nude mice and in conditions of interleukin-3 deprivation. To identify new miR-125b targets, we compared, by RNA-sequencing, the transcriptome of cell lines that do or do not over-express miR-125b.
Results: We showed that miR-125b over-expression blocks apoptosis and myeloid differentiation and enhances proliferation in both species. More importantly, we demonstrated that miR-125b is able to transform the 32Dclone3 cell line by conferring growth independence from interleukin-3; xenograft experiments showed that these cells form tumors in nude mice. Using RNA-sequencing and quantitative real-time polymerase chain reaction experiments, we identified multiple miR-125b targets. We demonstrated that ABTB1, an anti-proliferative factor, is a new direct target of miR-125b and we confirmed that CBFB, a transcription factor involved in hematopoiesis, is also targeted by miR-125b. MiR-125b controls apoptosis by down-regulating genes involved in the p53 pathway including BAK1 and TP53INP1.
Conclusions: This study demonstrates that in a myeloid context, miR-125b is an oncomiR able to transform cell lines. miR-125b blocks myeloid differentiation in part by targeting CBFB, blocks apoptosis through down-regulation of multiple genes involved in the p53 pathway, and confers a proliferative advantage to human and mouse myeloid cell lines in part by targeting ABTB1.Leukemia & Lymphoma Society of AmericaNational Institutes of Health (U.S.) (NIH grant DK068348)National Institutes of Health (U.S.) (NIH grant 5P01 HL066105
Balancing torques in membrane-mediated interactions: Exact results and numerical illustrations
Torques on interfaces can be described by a divergence-free tensor which is
fully encoded in the geometry. This tensor consists of two terms, one
originating in the couple of the stress, the other capturing an intrinsic
contribution due to curvature. In analogy to the description of forces in terms
of a stress tensor, the torque on a particle can be expressed as a line
integral along any contour surrounding the particle. Interactions between
particles mediated by a fluid membrane are studied within this framework. In
particular, torque balance places a strong constraint on the shape of the
membrane. Symmetric two-particle configurations admit simple analytical
expressions which are valid in the fully nonlinear regime; in particular, the
problem may be solved exactly in the case of two membrane-bound parallel
cylinders. This apparently simple system provides some flavor of the remarkably
subtle nonlinear behavior associated with membrane-mediated interactions.Comment: 16 pages, 10 figures, REVTeX4 style. The Gaussian curvature term was
included in the membrane Hamiltonian; section II.B was rephrased to smoothen
the flow of presentatio
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Corticotropinoma as a Component of Carney Complex.
Known germline gene abnormalities cause one-fifth of the pituitary adenomas in children and adolescents, but, in contrast with other pituitary tumor types, the genetic causes of corticotropinomas are largely unknown. In this study, we report a case of Cushing disease (CD) due to a loss-of-function mutation in PRKAR1A, providing evidence for association of this gene with a corticotropinoma. A 15-year-old male presenting with hypercortisolemia was diagnosed with CD. Remission was achieved after surgical resection of a corticotropin (ACTH)-producing pituitary microadenoma, but recurrence 3 years later prompted reoperation and radiotherapy. Five years after the original diagnosis, the patient developed ACTH-independent Cushing syndrome, and a diagnosis of primary pigmented nodular adrenocortical disease was confirmed. A PRKAR1A mutation (c.671delG, p.G225Afs*16) was detected in a germline DNA sample from the patient, which displayed loss of heterozygosity in the corticotropinoma. No other germline or somatic mutations of interest were found. As corticotropinomas are not a known component of Carney complex (CNC), we performed loss of heterozygosity and messenger RNA stability studies in the patient's tissues, and analyzed the effect of Prkar1a silencing on AtT-20/D16v-F2 mouse corticotropinoma cells. No PRKAR1A defects were found among 97 other pediatric CD patients studied. Our clinical case and experimental data support a role for PRKAR1A in the pathogenesis of a corticotroph cell tumor. This is a molecularly confirmed report of a corticotropinoma presenting in association with CNC. We conclude that germline PRKAR1A mutations are a novel, albeit apparently infrequent, cause of CD
Statistical Mechanics of Semiflexible Bundles of Wormlike Polymer Chains
We demonstrate that a semiflexible bundle of wormlike chains exhibits a
state-dependent bending stiffness that alters fundamentally its scaling
behavior with respect to the standard wormlike chain. We explore the
equilibrium conformational and mechanical behavior of wormlike bundles in
isolation, in crosslinked networks, and in solution.Comment: 4 pages, 3 figures, final version as publishe
How to determine local elastic properties of lipid bilayer membranes from atomic-force-microscope measurements: A theoretical analysis
Measurements with an atomic force microscope (AFM) offer a direct way to
probe elastic properties of lipid bilayer membranes locally: provided the
underlying stress-strain relation is known, material parameters such as surface
tension or bending rigidity may be deduced. In a recent experiment a
pore-spanning membrane was poked with an AFM tip, yielding a linear behavior of
the force-indentation curves. A theoretical model for this case is presented
here which describes these curves in the framework of Helfrich theory. The
linear behavior of the measurements is reproduced if one neglects the influence
of adhesion between tip and membrane. Including it via an adhesion balance
changes the situation significantly: force-distance curves cease to be linear,
hysteresis and nonzero detachment forces can show up. The characteristics of
this rich scenario are discussed in detail in this article.Comment: 14 pages, 9 figures, REVTeX4 style. New version corresponds to the
one accepted by PRE. The result section is restructured: a comparison to
experimental findings is included; the discussion on the influence of
adhesion between AFM tip and membrane is extende
Analytical study of the effect of recombination on evolution via DNA shuffling
We investigate a multi-locus evolutionary model which is based on the DNA
shuffling protocol widely applied in \textit{in vitro} directed evolution. This
model incorporates selection, recombination and point mutations. The simplicity
of the model allows us to obtain a full analytical treatment of both its
dynamical and equilibrium properties, for the case of an infinite population.
We also briefly discuss finite population size corrections
Loss-of-function mutations in the CABLES1 gene are a novel cause of Cushing's disease.
The CABLES1 cell cycle regulator participates in the adrenal-pituitary negative feedback, and its expression is reduced in corticotropinomas, pituitary tumors with a largely unexplained genetic basis. We investigated the presence of CABLES1 mutations/copy number variations (CNVs) and their associated clinical, histopathological and molecular features in patients with Cushing's disease (CD). Samples from 146 pediatric (118 germline DNA only/28 germline and tumor DNA) and 35 adult (tumor DNA) CD patients were screened for CABLES1 mutations. CNVs were assessed in 116 pediatric CD patients (87 germline DNA only/29 germline and tumor DNA). Four potentially pathogenic missense variants in CABLES1 were identified, two in young adults (c.532G > A, p.E178K and c.718C > T, p.L240F) and two in children (c.935G > A, p.G312D and c.1388A > G, and p.D463G) with CD; no CNVs were found. The four variants affected residues within or close to the predicted cyclin-dependent kinase-3 (CDK3)-binding region of the CABLES1 protein and impaired its ability to block cell growth in a mouse corticotropinoma cell line (AtT20/D16v-F2). The four patients had macroadenomas. We provide evidence for a role of CABLES1 as a novel pituitary tumor-predisposing gene. Its function might link two of the main molecular mechanisms altered in corticotropinomas: the cyclin-dependent kinase/cyclin group of cell cycle regulators and the epidermal growth factor receptor signaling pathway. Further studies are needed to assess the prevalence of CABLES1 mutations among patients with other types of pituitary adenomas and to elucidate the pituitary-specific functions of this gene
Cell-Free Synthesis of the Mitochondrial ADP/ATP Carrier Protein of Neurospora crassa
ADP/ATP carrier protein was synthesized in heterologous cell-free systems programmed with Neurospora poly(A)-containing RNA and homologous cell-free systems from Neurospora. The apparent molecular weight of the product obtained in vitro was the same as that of the authentic mitochondrial protein. The primary translation product obtained in reticulocyte lysates starts with formylmethionine when formylated initiator methionyl-tRNA (fMet-tRNAfMet) was present. The product synthesized in vitro was released from the ribosomes into the postribosomal supernatant.
The evidence presented indicates that the ADP/ATP carrier is synthesized as a polypeptide with the same molecular weight as the mature monomeric protein and does not carry an additional sequence
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