Rb Target Research Articles

Abstract 3450: ABT-348 in combination with inhibition of CDK4/6 highlights a strategy to target RB mutant cells while sparing RB wild-type cells.

Abstract ABT-348 is a novel, potent and orally bioavailable inhibitor of the Aurora kinases as well as the VEGF and PDGF families of receptor tyrosine kinases. ABT-348 is broadly efficacious preclinically as a single agent against a wide range of tumor types and is currently in Phase I/II clinical trials. Several published studies have supported the potential utility of combining targeted anti-proliferative agents with cytotoxic chemotherapies. Herein, the efficacy of combining the neocytotoxic agent, ABT-348, with CDK4/6 inhibition in the treatment of solid tumor cells has been evaluated. Inhibition of CDK4/6 via siRNA elicits a potent cytostatic response in cells that harbor a wild-type, functional RB. We demonstrate that features of cellular senescence are induced in human tumor cells with siRNAs targeting both CDK4 and CDK6. Dual inhibition of CDK4 and CDK6 is required for antiproliferative activity in most cancer cell lines and this is not accompanied by induction of apoptosis. In RB wild-type cells, CDK4/6 inhibition antagonizes the activity of ABT-348 by inducing G0/G1 arrest. Conversely, CDK4/6 inhibition does not alter the therapeutic response of RB-deficient cells to ABT-348, indicating that the effects of ABT-348 are dependent on cells progressing into mitosis. A preponderance of evidence from previous publications and our studies indicate that CDK4/6 inhibition attenuates the cellular response to cytotoxic chemotherapies in RB wild-type cells providing a potential to expand the therapeutic window for ABT-348. Thus combination with ABT-348 and a CDK4/6 inhibitor may provide an opportunity to selectively target RB mutant cells. Citation Format: Jun Guo, Michael L. Curtin, Zehan Chen, Daniel H. Albert, Paul Tapang, Yujia Dai, Mia-Ha Bui, Peter Kovar, Michael R. Michaelides, Keith B. Glaser, Chris Tse, O. Jameel Shah. ABT-348 in combination with inhibition of CDK4/6 highlights a strategy to target RB mutant cells while sparing RB wild-type cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3450. doi:10.1158/1538-7445.AM2013-3450

Inactivation of Rb in stromal fibroblasts promotes epithelial cell invasion.

Stromal-derived growth factors are required for normal epithelial growth but are also implicated in tumour progression. We have observed inactivation of the retinoblastoma protein (Rb), through phosphorylation, in cancer-associated fibroblasts in oro-pharyngeal cancer specimens. Rb is well known for its cell-autonomous effects on cancer initiation and progression; however, cell non-autonomous functions of Rb are not well described. We have identified a cell non-autonomous role of Rb, using three-dimensional cultures, where depletion of Rb in stromal fibroblasts enhances invasive potential of transformed epithelia. In part, this is mediated by upregulation of keratinocyte growth factor (KGF), which is produced by the depleted fibroblasts. KGF drives invasion of epithelial cells through induction of MMP1 expression in an AKT- and Ets2-dependent manner. Our data identify that stromal fibroblasts can alter the invasive behaviour of the epithelium, and we show that altered expression of KGF can mediate these functions.

P38α controls erythroblast enucleation and Rb signaling in stress erythropoiesis

Enucleation of erythroblasts during terminal differentiation is unique to mammals. Although erythroid enucleation has been extensively studied, only a few genes, including retinoblastoma protein (Rb), have been identified to regulate nuclear extrusion. It remains largely undefined by which signaling molecules, the extrinsic stimuli, such as erythropoietin (Epo), are transduced to induce enucleation. Here, we show that p38α, a mitogen-activated protein kinase (MAPK), is required for erythroid enucleation. In an ex vivo differentiation system that contains high Epo levels and mimics stress erythropoiesis, p38α is activated during erythroid differentiation. Loss of p38α completely blocks enucleation of primary erythroblasts. Moreover, p38α regulates erythroblast enucleation in a cell-autonomous manner in vivo during fetal and anemic stress erythropoiesis. Markedly, loss of p38α leads to downregulation of p21, and decreased activation of the p21 target Rb, both of which are important regulators of erythroblast enucleation. This study demonstrates that p38α is a key signaling molecule for erythroblast enucleation during stress erythropoiesis.

Abstract 2115: CASZ1 suppresses neuroblastoma cell growth by restoring Rb activity in G1 and regulating kinases involved in G2/M progression

Abstract A hallmark of cancer is the disruption of cell cycle regulation. Frequently this occurs in the Rb signal transduction pathway and causes tumor cells to become insensitive to antigrowth signals. In neuroblastoma (NB), poor prognosis tumors have been found to over-express multiple cell cycle regulatory genes, including those that participate in Rb regulation. Specifically, while amplification of Cyclin D1 and Cdk4 and mutations in Cdk6 occur at low frequency in NB, more than 75% of tumors contain elevated cyclin D1 expression. Recently, we identified CASZ1 to be a chromosome 1p36 tumor suppressor gene that suppresses NB tumor growth. To determine how loss of the CASZ1 tumor suppressor might contribute to cell cycle deregulation in NB tumors, we investigated how CASZ1 reconstitution affects proteins required for transition of NB cells through the G1/S and G2/M phases of the cell cycle by western blot analysis. CASZ1 restoration causes a 2-fold increase in the time required for NB cells to progress through the cell cycle with no major changes in any particular phase of the cell cycle. CASZ1 decreases DNA synthesis by ∼30% using a Click-iT EdU assay. Within 24 hr of CASZ1 induction there is a 2-fold increase in the level of the Cdk2/4 inhibitor p21 and a 1.5-fold increase in p27 levels at 72 hr. Steady-state levels of p16 levels do not change. There is also a 50% decrease in the steady-state levels of Cdk2 and Cdk6 at 24 hr. Consistent with the increase of Cdk inhibitors and decrease in Cdks levels, there is a 60% decrease in phosphorylation of their target Rb at p-RbS780 and p-RbS807/S811. Thus, increased levels of CASZ1 restore Rb activity, which regulates the G1/S transition. CASZ1 causes a near 40% decrease of cyclin E at 24 hr, and Cyclin E:Cdk2 is required for G1/S transition in addition to Rb. Cyclin A decreases about 40% at 24 hr after CASZ1 induction and cyclin A:Cdk2 is required for the cell to progress through the S phase. Cyclin B:Cdk1 is required for G2/M progression, CASZ1 induces 70% decrease of cyclin B1 and 40% decrease of its up-stream activator phosphates Cdc25c although Cdk1 level does not change. CASZ1 causes a 70% decrease in the steady-state levels of Aurora A kinase, which a key regulatory kinase in mitosis. These decreases occur at 48 hr suggesting that they may be related to changes occurring in cells progressing through G1. Taken together, this study indicates that restoration of CASZ1 activates Rb in G1 and inhibits the G2/M regulators cyclin B1, Cdc25c and aurora kinase. The decreases in the key regulators controlling G1/S and G2/M transition leads to a lengthening of NB cell cycle progression and a subsequent decreased growth in vitro and in vivo. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2115. doi:10.1158/1538-7445.AM2011-2115

Abstract 268: Differential regulation of MMP promoters by E2F transcription factors: Potential role of c-MYC and Id1

Abstract The retinoblastoma tumor suppressor, Rb, is the major negative regulator of E2F-1 transcription factor, and the Rb-E2F pathway is altered in most cancers. Further, many oncogenic mutations initiate tumors by targeting the Rb-E2F pathway. E2Fs regulate genes involved in a variety of biological processes, including angiogenesis, apoptosis, and differentiation, but it is unclear whether these molecules contribute to cancer cell invasion and metastasis. To investigate the role of E2F in metastasis, we analyzed the promoters of matrix metalloproteinase genes (MMPs), which are major contributors to the invasion and migration programs. We find that many MMP genes had potential E2F binding sites; we focused on MMP2, MMP9, MMP14, and MMP15 promoter, which have multiple E2F binding sites. Chip assays showed Rb, and E2Fs 1-5 could bind to the each MMP promoter. Transient transfection experiments with MMP promoter-luciferase constructs showed thatMMP9, MMP14, and MMP15-luc promoters were induced by E2Fs, whereas the MMP2 promoter was repressed by E2F1-5 in A549, and H1650 NSCLC cell lines. QRT-PCR showed that MMP2 was upregulated in A549 cells transfected with an siRNA targeting E2F1 and E2F3, whereas other MMPs were downregulated. To determine if MMP2 was repressed in an Rb-dependent manner, A549 cells stably expressing shRNA targeting Rb or a non-targeting control were transiently transfected with MMP2-luc construct and E2F1. In both cases, the MMP2-luc promoter was repressed by E2F1, suggesting that the repression is Rb-independent. Further, cells transfected with MMP2-luc promoter and E2F1 had less luciferase activity than cells transfected with MMP2 alone, independent of BRG-1, YY1, HDAC1, prohibitin-1, mSIN3a, or SUV39h1. Further, depletion of these co-repressors did not enhance MMP2 mRNA levels when compared to cells depleted of E2F1 alone. To determine if this repression is through binding site competition, we examined the effect of depleting transcription factors that have potential binding sites in the MMP2 promoter and were in close proximity to the E2F binding sites. We found that depletion of c-Myc and ID1 significantly enhanced MMP2 luciferase activity, and there was less repression from E2F1 in transient transfection experiments. These data suggest that E2F family members can repress the MMP2 promoter through mechanisms that may involve c-MYC and ID1. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 268. doi:10.1158/1538-7445.AM2011-268

Nuclear model calculation on charge particle induced reaction on Rb and Kr targets and targetry for 82Sr production

The 82Sr/82Rb radionuclide generator is used very commonly in positron emission tomography. ALICE/ASH and TALYS 1.0 codes were used to calculate excitation functions for proton, alpha and 3He induced on various targets that lead to produce 82Sr radioisotope using intermediate energy accelerators. Recommended thickness of the targets according to SRIM code was premeditated. The application of those data, particularly in the calculation of integral yields, is discussed and theoretical integral yields for any reaction were computed. To consider precision of TALYS 1.0 code calculations, 85Rb(p,4n)82Sr process was determined as most interesting one due to radionuclide purity. The TALYS 1.0 code predicts a maximum cross-section of about 130 mb at 47 MeV for this reaction. Rubidium chloride deposition on copper substrate was carried out via sedimentation method in order to produce 82Sr. 2.98 g RbCl, 1.043 g ethyl cellulose, 10 mL acetone were used to prepare a layer of enriched rubidium chloride of 11.69 cm2 area and 0.34 g/cm2 thickness.

RB-pathway disruption in breast cancer

In breast cancer, inactivation of the RB tumor suppressor gene is believed to occur via multiple mechanisms to facilitate tumorigenesis. However, the prognostic and predictive value of RB status in disease-specific clinical outcomes has remained uncertain. We investigated RB pathway deregulation in the context of both ER-positive and ER-negative disease using combined microarray datasets encompassing over 900 breast cancer patient samples. Disease-specific characteristics of RB pathway deregulation were investigated in this dataset by evaluating correlation among pathway genes as well as differential expression across patient tumor populations defined by ER status. Survival analysis among these breast cancer samples demonstrates that the RB-loss signature is associated with poor disease outcome within several independent cohorts. Within the ER-negative subpopulation, the RB-loss signature is associated with improved response to chemotherapy and longer relapse-free survival. Additionally, while individual genes in the RB target signature closely reproduce its prognostic value, they also serve to predict and monitor response to therapeutic compounds, such as the cytostatic agent PD-0332991. These results indicate that the RB-loss signature expression is associated with poor outcome in breast cancer, but predicts improved response to chemotherapy based on data in ER-negative populations. While the RB-loss signature, as a whole, demonstrates prognostic and predictive utility, a small subset of markers could be sufficient to stratify patients based on RB function and inform the selection of appropriate therapeutic regimens.

Targeting Rb Mutant Cancers by Inactivating TSC2

Retinoblastoma (Rb), a tumor suppressor gene, is inactivated in many types of cancer. However little is known about how the loss of Rb function can be targeted in cancer therapies. We have identified that inactivation of TSC2 in Rb mutant cancer cells will induce a synergistic cell death. The synergistic cell death is due to an increase in cellular stress including metabolic, ER, and oxidative stress. Therefore, inactivation of TSC2 and chemothereputics that result in induction of cellular stress may be a novel and effective way to treat cancers containing inactivated Rb.

Corrections: 2009 San Antonio Breast Cancer Symposium Abstracts

Corrections: 2009 San Antonio Breast Cancer Symposium Abstracts

A new MOTRIMS apparatus for high resolution measurements in ion-atom collisions and trapped atoms studies

A new MOTRIMS (magneto-optical trap-target recoil ion momentum spectroscopy) apparatus has been built at the LPC-CAEN, and recently tested with a beam of 2 keV Na+ projectiles colliding on a trapped Rb target. In this work, we demonstrate the capability of the setup to provide, with a very high signal over background ratio, fully differential cross sections in scattering angle, initial state, and final state of the system. The fine control of the trapping lasers, the extraction of the recoil ions transverse to the ion beam axis, and the fast switching for the MOT magnetic field, are three characteristics promising for new studies including collisions with oriented targets or requiring electron detection.

Inhibition of RNA Polymerase III Transcription by BRCA1

RNA polymerase III (RNA pol III) transcribes structural RNAs involved in RNA processing (U6 snRNA) and translation (tRNA), thereby regulating the growth rate of cells. Proper initiation by RNA pol III requires the transcription factor TFIIIB. Gene-external U6 snRNA transcription requires TFIIIB consisting of Bdp1, TBP, and Brf2. Transcription from the gene-internal tRNA promoter requires TFIIIB composed of Bdp1, TBP, and Brf1. TFIIIB is a target of tumor suppressors, including PTEN, ARF, p53, and RB, and RB-related pocket proteins. Breast cancer susceptibility gene 1 (BRCA1) tumor suppressor plays a role in DNA repair, cell cycle regulation, apoptosis, genome integrity, and ubiquitination. BRCA1 has a conserved amino-terminal RING domain, an activation domain 1 (AD1), and an acidic carboxyl-terminal domain (BRCA1 C-terminal region). In Saccharomyces cerevisiae, TFIIB interacts with the BRCA1 C-terminal region domain of Fcp1p, an RNA polymerase II phosphatase. The TFIIIB subunits Brf1 and Brf2 are structurally similar to TFIIB. Hence, we hypothesize that RNA pol III may be regulated by BRCA1 via the TFIIB family members Brf1 and Brf2. Here we report that: (1) BRCA1 inhibits both VAI (tRNA) and U6 snRNA RNA pol III transcription; (2) the AD1 of BRCA1 is responsible for inhibition of U6 snRNA transcription, whereas the RING domain and AD1 of BRCA1 are required for VAI transcription inhibition; and (3) overexpression of Brf1 and Brf2 alleviates inhibition of U6 snRNA and VAI transcription by BRCA1. Taken together, these data suggest that BRCA1 is a general repressor of RNA pol III transcription.

Regulation of the retinoblastoma proteins by the human herpesviruses

Viruses are obligate intracellular parasites that alter the environment of infected cells in order to replicate more efficiently. One way viruses achieve this is by modulating cell cycle progression. The main regulators of progression out of G0, through G1, and into S phase are the members of the retinoblastoma (Rb) family of tumor suppressors. Rb proteins repress the transcription of genes controlled by the E2F transcription factors. Because the expression of E2F-responsive genes is required for cell cycle progression into the S phase, Rb arrests the cell cycle in G0/G1. A number of viral proteins directly target Rb family members for inactivation, presumably to create an environment more hospitable for viral replication. Such viral proteins include the extensively studied oncoproteins E7 (from human papillomavirus), E1A (from adenovirus), and the large T (tumor) antigen (from simian virus 40). Elucidating how these three viral proteins target and inactivate Rb has proven to be an invaluable approach to augment our understanding of both normal cell cycle progression and carcinogenesis. In addition to these proteins, a number of other virally-encoded inactivators of the Rb family have subsequently been identified including a surprising number encoded by human herpesviruses. Here we review how the human herpesviruses modulate Rb function during infection, introduce the individual viral proteins that directly or indirectly target Rb, and speculate about what roles Rb modulation by these proteins may play in viral replication, pathogenesis, and oncogenesis.

Sorption of radiostrontium from liquid rubidium metal

Sorption of radiostrontium (in particular, 82Sr) from liquid rubidium metal on various inorganic sorbents (metals and oxides) was studied. The temperature dependence and dynamics of the adsorption were studied, and the observed relationships were accounted for. The procedure developed allows efficient isolation of 82Sr from Rb targets after their irradiation with accelerated protons. Various versions of the procedure are considered, and prospects for the development of a new high-performance process for preparing 82Sr, in particular, using circulating Rb targets, were discussed.

Human Papillomavirus E7 Requires the Protease Calpain to Degrade the Retinoblastoma Protein

Cervical cancers transformed by high risk human papilloma virus (HPV) express the E7 oncoprotein, which accelerates the degradation of the retinoblastoma protein (Rb). Here we show that the E7-mediated degradation of Rb requires the calcium-activated cysteine protease, calpain. E7 bound and activated mu-calpain and promoted cleavage at Rb(810), with mutation of this residue preventing E7-mediated degradation. The calpain cleavage product, Rb(1-810), was unable to mediate cell cycle arrest but retained the ability to repress E6/E7 transcription. E7 also promoted the accelerated proteasomal degradation of Rb(1-810). Calpain inhibitors reduced the viability of HPV-transformed cells and synergized with cisplatin. Calpain, thus, emerges as a central player in E7-mediated degradation of Rb and represents a potential new drug target for the treatment of HPV-associated lesions.

High mobility group protein HMGA1 inhibits retinoblastoma protein‐mediated cellular G0 arrest

Retinoblastoma protein (RB) acts as a tumor suppressor in many tissue types, by promoting cell arrest via E2F-mediated transcriptional repression. In addition to the aberrant forms of the RB gene found in different types of cancers, many viral oncoproteins including the simian virus 40 large T antigen target RB. However, cellular factors that inhibit RB function remain to be elucidated. Here, we report that RB interacts with the high mobility group protein A1 (HMGA1), a-non-histone architectural chromatin factor that is frequently overexpressed in cancer cells. HMGA1 binds the small pocket domain of RB, and competes with HDAC1. Subsequently, overexpression of HMGA1 abolishes the inhibitory effect of RB on E2F-activated transcription from the cyclin E promoter. Under serum starvation, T98G cells had been previously shown to be arrested in the G0 phase in an RB-mediated manner. The G0 phase was characterized by growth arrest and low levels of transcription, together with the hypophosphorylation of RB and the downregulation of HMGA1. In contrast, such serum-depleted G0 arrest was abrogated in T98G cells overexpressing HMGA1. The overexpressed HMGA1 was found to form complexes with cellular RB, suggesting that downregulation of HMGA1 is required for G0 arrest. There were no phenotypic changes in HMGA1-expressing T98G cells in the presence of serum, but the persistent expression of HMGA1 under serum starvation caused various nuclear abnormalities, which were similarly induced in T antigen-expressing T98G cells. Our present findings indicate that overexpression of HMGA1 disturbs RB-mediated cell arrest, suggesting a negative control of RB by HMGA1.

Electron capture cross-sections between 3He + ion and rubidium vapor in the energy range 1.0–19 keV

Electron capture cross-sections have been measured for 3He + ions in Rb vapor. The deduced data were used for evaluating the performance of the proposed polarized 3He ion source (SEPIS) based on the spin-exchange collisions. Measurements of the 3He + beam components after passing through the target were made as a function of the Rb target thickness at energies ranging from 1.0 to 19 keV. The Rb vapor thickness was varied by changing the Rb cell temperature and the thicknesses were calibrated by observing the Faraday rotation angles of a probe laser ( λ = 782.0 nm ) penetrating the Rb vapor under the presence of a magnetic field ( ∼ 0.26 T ) . Assuming that the double electron capture and stripping of 3He 0 atoms are neglected, the electron capture cross-sections σ c were deduced. It was found that the observed energy dependence of σ c significantly deviated from that of the He + 4 + Cs system which was measured in the past. Lastly, the observed energy dependence of σ c was compared with the theoretical calculation assuming the semi-classical close-coupling method based on the molecular-orbital expansion. It was found that the calculated results, as a whole, reproduced the experimental results except for the data collected at 1 keV. This qualitative reproduction partly supports the appropriateness of the above theory which predicts the large spin-exchange cross-sections at low energies.

Expression of Egr-1 gene and its correlation with the oncogene proteins in non-irradiated and irradiated esophageal squamous cell carcinoma

We study the expression of early growth response gene-1 (Egr-1 gene) in non-irradiated and irradiated human esophageal cancer tissues, and its relationship with the expression of C-fos, C-jun onco-proteins as well as Egr-1 target gene proteins P53, Rb and Bax expression. In situ hybridization (ISH) and immunohistochemistry (IHC) were used respectively to detect Egr-1 mRNA, Egr-1, C-fos, C-jun, P53, Rb and Bax proteins in 80 surgically resected non-irradiated and irradiated tumor specimens of esophageal squamous cell carcinoma. Egr-1 gene mRNA and Bax protein were located in the cytoplasm, whereas Egr-1, C-fos, C-jun, P53, Rb proteins were located in the nuclei. Egr-1 was expressed in nine out of 40 cases (22.5%) of non-irradiated and 23 of 40 cases (57.5%) of irradiated tumor specimens. No correlation was found between Egr-1 gene expression and C-fos, C-jun onco-proteins expression, neither was any correlation disclosed between Egr-1 gene expression with its target gene protein expression. Patients who underwent radiotherapy with Egr-1 overexpressed in their cancer tissue had better prognosis. Radiotherapy up-regulates Egr-1 expression in esophageal carcinoma. Egr-1 overexpression may be a potential radiation response gene marker and may play an important role in prognosis of esophageal squamous cell carcinoma.

Over-expressed estrogen receptor-α up-regulates hTNF-α gene expression and down-regulates β-catenin signaling activity to induce the apoptosis and inhibit proliferation of LoVo colon cancer cells

Epidemiologic studies reported that the prevalence of hereditary non-polyposis colon cancer (HNPCC) in male is about 1.5-fold higher than that in female. Decreases in circulatory estrogen (E(2)) have been reported to downregulate the expression of E(2) receptor (ER) and significantly increase the risk of colorectal cancer. Patients that received E(2) replacement therapy were found to have a reduction in the incidence of colon adenoma and carcinoma. Furthermore, significant decreases in the expression of ER have been found in colorectal cancer specimens. Evidences strongly suggest the protective roles of E(2) and ER against colorectal cancer. However, the mechanisms of ERalpha effects on colorectal cancer cells remained un-clear. LoVo cells were transient transfected to overexpress ERalpha, DNA fragmentation and the activated caspases measurements were performed to evaluate apoptotic effects. Western blotting was used to evaluate protein levels, and luciferase activity assay to measure the Htnf-a promoter activity. The results clearly demonstrated that overexpressed ERalpha with or without E(2) (10(-8) M) treatment could activate caspase -8, -9, and 3 and induce DNA fragmentation in LoVo cell. At the same time, overexpressed ERalpha plus E(2) significantly increases the expression and promoter activity of hTNF-alpha, and the DNA fragmentation effect induced by E(2) plus ERalpha were reduced by the addition of hTNF antibody (0.1 ng(ml). In addition, E(2) plus ERalpha significantly upregulated p21 and p27 levels and downregulated the beta-catenin and its target genes, cyclin D1 and Rb, which regulate the cell cycle and cell proliferation. The results indicate that E(2) plus overexpressed ERalpha induce LoVo cell apoptosis might mediate through the increase of hTNF-alpha gene expression, which in turn activate caspase-8, -9 and caspase-3 and lead to the DNA fragmentation and apoptosis. E(2) plus ERalpha also showed the downregulation of beta-catenin signalings implicating the suppression of proliferation and metastasis of colorectal cells. Efforts aiming at enhancing ERalpha expression and(or activity may be proved to be an alternative therapy against colorectal cancer.

S-wave elastic scattering of antihydrogen off atomic alkali-metal targets

We have investigated the $s$-wave elastic scattering of antihydrogen atoms off atomic alkali-metal targets (Li, Na, K, and Rb) at thermal energies $({10}^{\ensuremath{-}16}--{10}^{\ensuremath{-}4}\phantom{\rule{0.3em}{0ex}}\mathrm{a.u.})$ using an atomic orbital expansion technique. The elastic cross sections of these systems at thermal energies are found to be very high compared to $\overline{\mathrm{H}}\text{\ensuremath{-}}\mathrm{H}$ and $\overline{\mathrm{H}}\text{\ensuremath{-}}\mathrm{He}$ systems. The theoretical models employed in this study are so chosen to consider long-range forces dynamically in the calculation. The mechanism of cooling suggests that Li may be considered to be a good candidate as a buffer gas for enhanced cooling of antihydrogen atoms to ultracold temperature.

Epstein–Barr virus latent antigen 3C can mediate the degradation of the retinoblastoma protein through an SCF cellular ubiquitin ligase

Epstein-Barr virus (EBV) stimulates the proliferation of latently infected B cells and promotes lymphoid malignancies in humans. To address the role of EBV latency protein Epstein-Barr nuclear antigen 3C (EBNA3C) in regulation of the retinoblastoma protein (Rb), we transfected EBNA3C into 293, BJAB, and SAOS-2 cells. In this context, a dominant effect of EBNA3C is to decrease Rb protein levels. EBNA3C also rescues an Rb-induced flat cell phenotype and targets Rb for proteasome- and ubiquitin-dependent degradation. Further, EBNA3C forms a stable complex with Rb in cells when the proteasome machinery is inhibited and interacts with Rb in vitro, mapping to a conserved domain at the terminus of EBNA3C. Deletion analysis of EBNA3C identified a motif within amino acids 140-149 important for both the binding and regulation of Rb. This motif is of particular interest, because it has also been linked to regulation of the Skp1/Cul1/F-box complex, SCF(Skp2). Indeed, inhibition of Skp2 function with a dominant-negative molecule reduces the ability of EBNA3C to degrade Rb. Skp2 has no detectable effect on Rb levels in the absence of EBNA3C, suggesting that SCF(Skp2) is specifically usurped by EBNA3C for the enhancement of Rb degradation. That EBNA3C has exploited this association suggests that other human malignancies might use a similar strategy to regulate the Rb protein.