Medicine and Health Sciences Commons^™

Clarithromycin Attenuates Il-13–Induced Periostin Production In Human Lung Fibroblasts, Kosaku Komiya, Shoichiro Ohta, Kazuhiko Arima, Masahiro Ogawa, Shoichi Suzuki, Yasutaka Mitamura, Satoshi Nunomura, Yasuhiro Nanri, Tomohito Yoshihara, Atsushi Kawaguchi, Jun-Ichi Kadota, Bruce K. Rubin, Kenji Izuhara

Biochemistry and Molecular Biology Publications

Background

Periostin is a biomarker indicating the presence of type 2 inflammation and submucosal fibrosis; serum periostin levels have been associated with asthma severity. Macrolides have immunomodulatory effects and are considered a potential therapy for patients with severe asthma. Therefore, we investigated whether macrolides can also modulate pulmonary periostin production.

Methods

Using quantitative PCR and ELISA, we measured periostin production in human lung fibroblasts stimulated by interleukin-13 (IL-13) in the presence of two 14-member–ring macrolides—clarithromycin or erythromycin—or a 16-member–ring macrolide, josamycin. Phosphorylation of signal transducers and activators of transcription 6 (STAT6), downstream of IL-13 signaling, was evaluated by Western blotting. …

Neutralization Of Diverse Human Cytomegalovirus Strains Conferred By Antibodies Targeting Viral Gh/Gl/Pul128-131 Pentameric Complex, Sha Ha, Fengsheng Li, Matthew C. Troutman, Daniel C. Freed, Aimin Tang, John W. Loughney, Dai Wang, I-Ming Wang, Josef Vlasak, David C. Nickle, Richard R. Rustandi, Melissa Hamm, Pete A. Dephillips, Ningyan Zhang, Jason S. Mclellan, Hua Zhu, Stuart P. Adler, Michael A. Mcvoy, Zhiqiang An, Tong-Ming Fu

Biochemistry and Molecular Biology Publications

Human cytomegalovirus (HCMV) is the leading cause of congenital viral infection, and developing a prophylactic vaccine is of high priority to public health. We recently reported a replication-defective human cytomegalovirus with restored pentameric complex glycoprotein H (gH)/gL/pUL128-131 for prevention of congenital HCMV infection. While the quantity of vaccine-induced antibody responses can be measured in a viral neutralization assay, assessing the quality of such responses, including the ability of vaccine-induced antibodies to cross-neutralize the field strains of HCMV, remains a challenge. In this study, with a panel of neutralizing antibodies from three healthy human donors with natural HCMV infection or a …

Directed Evolution Provides Insight Into Conformational Substrate Sampling By Srta, Muna Suliman, Vishaka Santosh, Tom C. M. Seegar, Annamarie C. Dalton, Kathryn M. Schultz, Candice S. Klug, William A. Barton

Biochemistry and Molecular Biology Publications

The Sortase family of transpeptidases are found in numerous gram-positive bacteria and involved in divergent physiological processes including anchoring of surface proteins to the cell wall as well as pili assembly. As essential proteins, sortase enzymes have been the focus of considerable interest for the development of novel anti-microbials, however, more recently their function as unique transpeptidases has been exploited for the synthesis of novel bio-conjugates. Yet, for synthetic purposes, SrtA-mediated conjugation suffers from the enzyme's inherently poor catalytic efficiency. Therefore, to identify SrtA variants with improved catalytic efficiency, we used directed evolution to select a catalytically enhanced SrtA enzyme. …

Slowly Cycling Rho Kinase-Dependent Actomyosin Cross-Bridge "Slippage" Explains Intrinsic High Compliance Of Detrusor Smooth Muscle, Christopher J. Neal, Jia B. Lin, Tanner Hurley, Amy S. Miner, John E. Speich, Adam P. Klausner, Paul H. Ratz

Biochemistry and Molecular Biology Publications

Biological soft tissues are viscoelastic because they display timeindependent pseudoelasticity and time-dependent viscosity. However, there is evidence that the bladder may also display plasticity, defined as an increase in strain that is unrecoverable unless work is done by the muscle. In the present study, an electronic lever was used to induce controlled changes in stress and strain to determine whether rabbit detrusor smooth muscle (rDSM) is best described as viscoelastic or viscoelastic plastic. Using sequential ramp loading and unloading cycles, stress-strain and stiffness-stress analyses revealed that rDSM displayed reversible viscoelasticity, and that the viscous component was responsible for establishing a …

Cryptosporidium Priming Is More Effective Than Vaccine For Protection Against Cryptosporidiosis In A Murine Protein Malnutrition Model, Luther A. Bartelt, David T. Bolick, Glynis L. Kolling, James K. Roche, Edna I. Zaenker, Ana M. Lara, Francisco Jose Noronha, Carrie A. Cowardin, John H. Moore, Jerrold R. Turner, Cirle A. Warren, Gregory A. Buck, Richard L. Guerrant

Biochemistry and Molecular Biology Publications

Cryptosporidium is a major cause of severe diarrhea, especially in malnourished children. Using a murine model of C. parvum oocyst challenge that recapitulates clinical features of severe cryptosporidiosis during malnutrition, we interrogated the effect of protein malnutrition (PM) on primary and secondary responses to C. parvum challenge, and tested the differential ability of mucosal priming strategies to overcome the PM-induced susceptibility. We determined that while PM fundamentally alters systemic and mucosal primary immune responses to Cryptosporidium, priming with C. parvum (106 oocysts) provides robust protective immunity against re-challenge despite ongoing PM. C. parvum priming restores mucosal Th1-type effectors (CD3+CD8+CD103+ T-cells) …

Constitutive Association Of Tie1 And Tie2 With Endothelial Integrins Is Functionally Modulated By Angiopoietin-1 And Fibronectin, Annamarie C. Dalton, Tomer Shlamkovitch, Niv Papo, William A. Barton

Biochemistry and Molecular Biology Publications

Functional cross-talk between Tie2 and Integrin signaling pathways is essential to coordinate endothelial cell adhesion and migration in response to the extracellular matrix, yet the mechanisms behind this phenomenon are unclear. Here, we examine the possibility that receptor cross-talk is driven through uncharacterized Tie-integrin interactions on the endothelial surface. Using a live cell FRET-based proximity assay, we monitor Tie-integrin receptor recognition and demonstrate that both Tie1 and Tie2 readily associate with integrins α5ß1 and αVß3 through their respective ectodomains. Although not required, Tie2-integrin association is significantly enhanced in the presence of the extracellular component and integrin ligand fibronectin. In vitro …

Mir-9 Acts As An Oncomir In Prostate Cancer Through Multiple Pathways That Drive Tumour Progression And Metastasis, S. J. Seashols-Williams, W. Budd, G. C. Clark, Q. Wu, R. Daniel, E. Dragoescu, Z. E. Zehner

Biochemistry and Molecular Biology Publications

Identification of dysregulated microRNAs (miRNAs) in prostate cancer is critical not only for diagnosis, but also differentiation between the aggressive and indolent forms of the disease. miR-9 was identified as an oncomiR through both miRNA panel RT-qPCR as well as high-throughput sequencing analysis of the human P69 prostate cell line as compared to its highly tumorigenic and metastatic subline M12, and found to be consistently upregulated in other prostate cell lines including DU-145 and PC3. While miR-9 has been characterized as dysregulated either as an oncomiR or tumour suppressor in a variety of other cancers including breast, ovarian, and nasopharyngeal …

Lysophosphatidic Acid Up-Regulates Hexokinase Ii And Glycolysis To Promote Proliferation Of Ovarian Cancer Cells, Abir Mukherjee, Yibao Ma, Fang Yuan, Yongling Gong, Zhenyu Fang, Esraa M. Mohamed, Erika Berrios, Huanjie Shao, Xianjun Fang

Biochemistry and Molecular Biology Publications

Lysophosphatidic acid (LPA), a blood-borne lipid mediator, is present in elevated concentrations in ascites of ovarian cancer patients and other malignant effusions. LPA is a potent mitogen in cancer cells. The mechanism linking LPA signal to cancer cell proliferation is not well understood. Little is known about whether LPA affects glucose metabolism to accommodate rapid proliferation of cancer cells. Here we describe that in ovarian cancer cells, LPA enhances glycolytic rate and lactate efflux. A real time PCR-based miniarray showed that hexokinase II (HK2) was the most dramatically induced glycolytic gene to promote glycolysis in LPA-treated cells. Analysis of the …

Dual Action Of Mir-125b As A Tumor Suppressor And Oncomir-22 Promotes Prostate Cancer Tumorigenesis, William T. Budd, Sarah J. Seashols-Williams, Gene C. Clark, Danielle Weaver, Valerie Calvert, Emanuel Petricoin, Ema A. Dragoescu, Katherine O'Hanlon, Zendra E. Zehner

Biochemistry and Molecular Biology Publications

MicroRNAs (miRs) are a novel class of small RNA molecules, the dysregulation of which can contribute to cancer. A combinatorial approach was used to identify miRs that promote prostate cancer progression in a unique set of prostate cancer cell lines, which originate from the parental p69 cell line and extend to a highly tumorigenic/metastatic M12 subline. Together, these cell lines are thought to mimic prostate cancer progression in vivo. Previous network analysis and miR arrays suggested that the loss of hsa-miR-125b together with the overexpression of hsa-miR-22 could contribute to prostate tumorigenesis. The dysregulation of these two miRs was …

Dpy-17 And Mua-3 Interact For Connective Tissue-Like Tissue Integrity In Caenorhabditis Elegans: A Model For Marfan Syndrome, Pauline Fotopoulos, Jeongho Kim, Moonjung Hyun, Waiss Qamari, Inhwan Lee, Young-Jai You

Biochemistry and Molecular Biology Publications

mua-3 is a Caenorhabditis elegans homolog of the mammalian fibrillin1, a monogenic cause of Marfan syndrome. We identified a new mutation of mua-3 that carries an in-frame deletion of 131 amino acids in the extracellular domain, which allows the mutants to survive in a temperature-dependent manner; at the permissive temperature, the mutants grow normally without obvious phenotypes, but at the nonpermissive temperature, more than 90% die during the L4 molt due to internal organ detachment. Using the temperature-sensitive lethality, we performed unbiased genetic screens to isolate suppressors to find genetic interactors of MUA-3. From two independent screens, we isolated …

Dissection Of C. Elegans Behavioral Genetics In 3-D Environments, Namseop Kwon, Ara B. Hwang, Young-Jai You, Seung-Jae V. Lee, Jung Ho Je

Biochemistry and Molecular Biology Publications

The nematode Caenorhabditis elegans is a widely used model for genetic dissection of animal behaviors. Despite extensive technical advances in imaging methods, it remains challenging to visualize and quantify C. elegans behaviors in three-dimensional (3-D) natural environments. Here we developed an innovative 3-D imaging method that enables quantification of C. elegans behavior in 3-D environments. Furthermore, for the first time, we characterized 3-D-specific behavioral phenotypes of mutant worms that have defects in head movement or mechanosensation. This approach allowed us to reveal previously unknown functions of genes in behavioral regulation. We expect that our 3-D imaging method will facilitate new …

Missed Opportunities For The Diagnosis Of Colorectal Cancer, Laura A. Siminoff, Heather L. Rogers, Sonja Harris-Haywood

Biochemistry and Molecular Biology Publications

Objective. To examine patient and medical characteristics which predict a missed diagnostic opportunity (MDO) for colorectal cancer (CRC). Methods. The sample consisted of 252 patients diagnosed with Stages 1–4 CRC who were diagnosed in the prior six months, had experienced symptoms prior to diagnosis, and were not diagnosed through routine screening. Systematic review of all medical records prior to patients’ diagnosis was conducted. An MDO was defined as a clinical encounter where, even in the presence of presumptive CRC symptoms, the CRC diagnostic process is not started. Results. 92 patients (36.5%) experienced an MDO. Almost 80% of alternate diagnoses were …

Network Modelling Reveals The Mechanism Underlying Colitis-Associated Colon Cancer And Identifies Novel Combinatorial Anti-Cancer Targets, Junyan Lu, Hanlin Zeng, Zhongjie Liang, Limin Chen, Liyi Zhang, Hao Zhang, Hong Liu, Hualiang Jiang, Bairong Shen, Ming Huang, Meiyu Geng, Sarah Spiegel, Cheng Luo

Biochemistry and Molecular Biology Publications

The connection between inflammation and tumourigenesis has been well established. However, the detailed molecular mechanism underlying inflammation-associated tumourigenesis remains unknown because this process involves a complex interplay between immune microenvironments and epithelial cells. To obtain a more systematic understanding of inflammation-associated tumourigenesis as well as to identify novel therapeutic approaches, we constructed a knowledge-based network describing the development of colitis-associated colon cancer (CAC) by integrating the extracellular microenvironment and intracellular signalling pathways. Dynamic simulations of the CAC network revealed a core network module, including P53, MDM2, and AKT, that may govern the malignant transformation of colon epithelial cells in a …

P. Falciparum And P. Vivax Epitope-Focused Vlps Elicit Sterile Immunity To Blood Stage Infections, David C. Whitacre, Diego A. Espinosa, Cory J. Peters, Joyce E. Jones, Amy E. Tucker, Darrell L. Peterson, Fidel P. Zavala, David R. Milich

Biochemistry and Molecular Biology Publications

In order to design P. falciparum preerythrocytic vaccine candidates, a library of circumsporozoite (CS) T and B cell epitopes displayed on the woodchuck hepatitis virus core antigen (WHcAg) VLP platform was produced. To test the protective efficacy of the WHcAg-CS VLPs, hybrid CS P. berghei/P. falciparum (Pb/Pf) sporozoites were used to challenge immunized mice. VLPs carrying 1 or 2 different CS repeat B cell epitopes and 3 VLPs carrying different CS non-repeat B cell epitopes elicited high levels of anti-insert antibodies (Abs). Whereas, VLPs carrying CS repeat B cell epitopes conferred 98% protection of the liver against …

Interrogating The Venom Of The Viperid Snake Sistrurus Catenatus Edwardsii By A Combined Approach Of Electrospray And Maldi Mass Spectrometry, Alex Chapeaurouge, Md Abu Reza, Stephen P. Mackessy, Paulo C. Carvalho, Richard H. Valente, André Teixeira-Ferreira, Jonas Perales, Qingsong Lin, R. Manjunatha Kini

Biochemistry and Molecular Biology Publications

The complete sequence characterization of snake venom proteins by mass spectrometry is rather challenging due to the presence of multiple isoforms from different protein families. In the present study, we investigated the tryptic digest of the venom of the viperid snake Sistrurus catenatus edwardsii by a combined approach of liquid chromatography coupled to either electrospray (online) or MALDI (offline) mass spectrometry. These different ionization techniques proved to be complementary allowing the identification a great variety of isoforms of diverse snake venom protein families, as evidenced by the detection of the corresponding unique peptides. For example, ten out of eleven predicted …

Increased Eicosanoid Levels In The Sugen/Chronic Hypoxia Model Of Severe Pulmonary Hypertension, Aysar Al-Husseini, Dayanjan S. Wijesinghe, Laszlo Farkas, Donatas Kraskauskas, Jennifer I. Drake, Ben Van Tassel, Antonio Abbate, Charles E. Chalfant, Norbert F. Voelkel

Biochemistry and Molecular Biology Publications

Inflammation and altered immunity are recognized components of severe pulmonary arterial hypertension in human patients and in animal models of PAH. While eicosanoid metabolites of cyclooxygenase and lipoxygenase pathways have been identified in the lungs from pulmonary hypertensive animals their role in the pathogenesis of severe angioobliterative PAH has not been examined. Here we investigated whether a cyclooxygenase-2 (COX-2) inhibitor or diethylcarbamazine (DEC), that is known for its 5-lipoxygenase inhibiting and antioxidant actions, modify the development of PAH in the Sugen 5416/hypoxia (SuHx) rat model. The COX-2 inhibitor SC-58125 had little effect on the right ventricular pressure and did not …

Dual Action Of Mir-125b As A Tumor Suppressor And Oncomir-22 Promotes Prostate Cancer Tumorigenesis, William Budd, Sarah J. Seashols-Williams, Gene C. Clark, Danielle Weaver, Valerie Calvert, Emanuel Petricoin, Ema A. Dragoescu, Katherine O’Hanlon, Zendra E. Zehner

Biochemistry and Molecular Biology Publications

MicroRNAs (miRs) are a novel class of small RNA molecules, the dysregulation of which can contribute to cancer. A combinatorial approach was used to identify miRs that promote prostate cancer progression in a unique set of prostate cancer cell lines, which originate from the parental p69 cell line and extend to a highly tumorigenic/metastatic M12 subline. Together, these cell lines are thought to mimic prostate cancer progression in vivo. Previous network analysis and miR arrays suggested that the loss of hsa-miR-125b together with the overexpression of hsa-miR-22 could contribute to prostate tumorigenesis. The dysregulation of these two miRs was …

Pde5 Inhibitors Enhance Celecoxib Killing In Multiple Tumor Types, Laurence Booth, Jane L. Roberts, Nichola Cruickshanks, Seyedmehrad Tavallai, Timothy Webb, Peter Samuel, Adam Conley, Brittany Binion, Harold F. Young, Andrew Poklepovic, Sarah Spiegel, Paul Dent

Biochemistry and Molecular Biology Publications

The present studies determined whether clinically relevant phosphodiesterase 5 (PDE5) inhibitors interacted with a clinically relevant NSAID, celecoxib, to kill tumor cells. Celecoxib and PDE5 inhibitors interacted in a greater than additive fashion to kill multiple tumor cell types. Celecoxib and sildenafil killed ex vivo primary human glioma cells as well as their associated activated microglia. Knock down of PDE5 recapitulated the effects of PDE5 inhibitor treatment; the nitric oxide synthase inhibitor L-NAME suppressed drug combination toxicity. The effects of celecoxib were COX2 independent. Over-expression of c-FLIP-s or knock down of CD95/FADD significantly reduced killing by the drug combination. CD95 …

Dpy-17 And Mua-3 Interact For Connective Tissue-Like Tissue Integrity In Caenorhabditis Elegans: A Model For Marfan Syndrome, Pauline Fotopoulos, Jeongho Kim, Moonjung Hyun, Waiss Qamari, Inhwan Lee, Young-Jai You

Biochemistry and Molecular Biology Publications

mua-3 is a Caenorhabditis elegans homolog of the mammalian fibrillin1, a monogenic cause of Marfan syndrome. We identified a new mutation of mua-3that carries an in-frame deletion of 131 amino acids in the extracellular domain, which allows the mutants to survive in a temperature-dependent manner; at the permissive temperature, the mutants grow normally without obvious phenotypes, but at the nonpermissive temperature, more than 90% die during the L4 molt due to internal organ detachment. Using the temperature-sensitive lethality, we performed unbiased genetic screens to isolate suppressors to find genetic interactors of MUA-3. From two independent screens, we isolated …

The Signal Transducer And Activator Of Transcription 1 (Stat1) Inhibits Mitochondrial Biogenesis In Liver And Fatty Acid Oxidation In Adipocytes, Jennifer D. Sisler, Magdalena Morgan, Vidisha Raje, Rebecca C. Grande, Marta Derecka, Jeremy Meier, Marc Cantwell, Karol Szczepanek, William J. Korzun, Edward J. Lesnefsky, Thurl E. Harris, Colleen M. Croniger, Andrew C. Larner

Biochemistry and Molecular Biology Publications

The transcription factor STAT1 plays a central role in orchestrating responses to various pathogens by activating the transcription of nuclear-encoded genes that mediate the antiviral, the antigrowth, and immune surveillance effects of interferons and other cytokines. In addition to regulating gene expression, we report that STAT1-/- mice display increased energy expenditure and paradoxically decreased release of triglycerides from white adipose tissue (WAT). Liver mitochondria from STAT1-/- mice show both defects in coupling of the electron transport chain (ETC) and increased numbers of mitochondria. Consistent with elevated numbers of mitochondria, STAT1-/- mice expressed increased amounts of PGC1α, a …

The Phosphorylated Prodrug Fty720 Is A Histone Deacetylase Inhibitor That Reactivates Erα Expression And Enhances Hormonal Therapy For Breast Cancer, Nitai C. Hait, Dorit Avni, Akimitsu Yamada, Masayuki Nagahashi, Tomoyoshi Aoyagi, Hiroaki Aoki, Catherine I. Dumur, Z Zelenko, E J. Gallagher, D Leroith, Sheldon Milstien, Kazuaki Takabe, Sarah Spiegel

Biochemistry and Molecular Biology Publications

Estrogen receptor-α (ERα)-negative breast cancer is clinically aggressive and does not respond to conventional hormonal therapies. Strategies that lead to re-expression of ERα could sensitize ERα-negative breast cancers to selective ER modulators. FTY720 (fingolimod, Gilenya), a sphingosine analog, is the Food and Drug Administration (FDA)-approved prodrug for treatment of multiple sclerosis that also has anticancer actions that are not yet well understood. We found that FTY720 is phosphorylated in breast cancer cells by nuclear sphingosine kinase 2 and accumulates there. Nuclear FTY720-P is a potent inhibitor of class I histone deacetylases (HDACs) that enhances histone acetylations and regulates expression of …

Memo Has A Novel Role In S1p Signaling And Crucial For Vascular Development, Shunya Kondo, Alessia Bottos, Jeremy Allegood, Regis Masson, Francisca G. Maurer, Christel Genoud, Patrick Kaeser, Andrea Huwiler, Masato Murakami, Sarah Spiegel, Nancy E. Hynes

Biochemistry and Molecular Biology Publications

Memo is a conserved protein that was identified as an essential mediator of tumor cell motility induced by receptor tyrosine kinase activation. Here we show that Memo null mouse embryonic fibroblasts (MEFs) are impaired in PDGF-induced migration and this is due to a defect in sphingosine-1-phosphate (S1P) signaling. S1P is a bioactive phospholipid produced in response to multiple stimuli, which regulates many cellular processes. S1P is secreted to the extracellular milieu where it exerts its function by binding a family of G-protein coupled receptors (S1PRs), causing their activation in an autocrine or paracrine manner. The process, termed cell-autonomous S1PR signaling, …

Otitis Media In Sperm-Associated Antigen 6 (Spag6)-Deficient Mice, Xiaofei Li, Lei Xu, Jianfeng Li, Boqin Li, Xiaohui Bai, Jerome F. Strauss Iii, Zhibing Zhang, Haibo Wang

Biochemistry and Molecular Biology Publications

Mammalian SPAG6 protein is localized to the axoneme central apparatus, and it is required for normal flagella and cilia motility. Recent studies demonstrated that the protein also regulates ciliogenesis and cilia polarity in the epithelial cells of brain ventricles and trachea. Motile cilia are also present in the epithelial cells of the middle ear and Eustachian tubes, where the ciliary system participates in the movement of serous fluid and mucus in the middle ear. Cilia defects are associated with otitis media (OM), presumably due to an inability to efficiently transport fluid, mucus and particles including microorganisms. We investigated the potential …

Structure-Based Mechanism For Early Plp-Mediated Steps Of Rabbit Cytosolic Serine Hydroxymethyltransferase Reaction, Martino L. Di Salvo, J. Neel Scarsdale, Galina Kazanina, Roberto Contestabile, Verne Schirch, H. Tonie Wright

Biochemistry and Molecular Biology Publications

Serine hydroxymethyltransferase catalyzes the reversible interconversion of L-serine and glycine with transfer of one-carbon groups to and from tetrahydrofolate. Active site residue Thr254 is known to be involved in the transaldimination reaction, a crucial step in the catalytic mechanism of all pyridoxal 5′-phosphate- (PLP-) dependent enzymes, which determines binding of substrates and release of products. In order to better understand the role of Thr254, we have expressed, characterized, and determined the crystal structures of rabbit cytosolic serine hydroxymethyltransferase T254A and T254C mutant forms, in the absence and presence of substrates. These mutants accumulate a kinetically stable gem-diamine intermediate, and their …

Osu-03012 Sensitizes Breast Cancers To Lapatinib-Induced Cell Killing: A Role For Nck1 But Not Nck2, N. Winston West, Aileen Garcia-Vargas, Charles E. Chalfant, Margaret A. Park

Biochemistry and Molecular Biology Publications

Background

Lapatinib is characterized as an ErbB1/ErbB2 dual inhibitor and has recently been approved for the treatment of metastatic breast cancer. In this study, we examined mechanisms associated with enhancing the activity of lapatinib via combination with other therapies.

Methods

In the present studies, estrogen receptor (ER) positive and ER negative breast cancer cells were genetically manipulated to up- or downregulate eIF2-alpha, its phospho-mutant, Nck1, or Nck2, then treated with OSU-03012, lapatinib or the combination and assayed for cytotoxicity/cytostaticity using clonogenic assays.

Results

Treatment of breast cancer cell lines with lapatinib and OSU-03012 (a small molecule derivative of the Cox-2 …

A Networks Method For Ranking Microrna Dysregulation In Cancer, William Budd, Sarah Seashols, Danielle Weaver, Cyriac Joseph, Zendra E. Zehner

Biochemistry and Molecular Biology Publications

Background

Despite the lack of agreement on their exact roles, it is known that miRNAs contribute to cancer progression. Many studies utilize methods to detect differential regulation of miRNA expression. It is prohibitively expensive to examine all potentially dysregulated miRNAs and traditionally, researchers have focused their efforts on the most extremely dysregulated miRNAs. These methods may overlook the contribution of less differentially expressed but more functionally relevant miRNAs. The purpose of this study was to outline a method that not only utilizes differential expression but ranks miRNAs based on the functional relevance of their targets. This work uses a networks …

The Geometry Of Locomotive Behavioral States In C. Elegans, Thomas Gallagher, Theresa Bjorness, Robert Greene, Young-Jai You, Leon Avery

Biochemistry and Molecular Biology Publications

We develop a new hidden Markov model-based method to analyze C elegans locomotive behavior and use this method to quantitatively characterize behavioral states. In agreement with previous work, we find states corresponding to roaming, dwelling, and quiescence. However, we also find evidence for a continuum of intermediate states. We suggest that roaming, dwelling, and quiescence may best be thought of as extremes which, mixed in any proportion, define the locomotive repertoire of C elegans foraging and feeding behavior.

Light-Induced Translocation Of Rgs9-1 And Gβ5l In Mouse Rod Photoreceptors, Mei Tian, Marisa Zallocchi, Weimin Wang, Ching-Kang Chen, Krzysztof Palczewski, Duane Delimont, Dominic Cosgrove, You-Wei Peng

Biochemistry and Molecular Biology Publications

The transducin GTPase-accelerating protein complex, which determines the photoresponse duration of photoreceptors, is composed of RGS9-1, Gβ5L and R9AP. Here we report that RGS9-1 and Gβ5L change their distribution in rods during light/dark adaptation. Upon prolonged dark adaptation, RGS9-1 and Gβ5L are primarily located in rod inner segments. But very dim-light exposure quickly translocates them to the outer segments. In contrast, their anchor protein R9AP remains in the outer segment at all times. In the dark, Gβ5L's interaction with R9AP decreases significantly and RGS9-1 is phosphorylated at S475 to a significant degree. Dim light exposure leads to quick de-phosphorylation of …

Hif- And Non-Hif-Regulated Hypoxic Responses Require The Estrogen-Related Receptor In Drosophila Melanogaster, Yan Li, Divya Padmanabha, Luciana B. Gentile, Catherine I. Dumur, Robert B. Beckstead, Keith D. Baker

Biochemistry and Molecular Biology Publications

Low-oxygen tolerance is supported by an adaptive response that includes a coordinate shift in metabolism and the activation of a transcriptional program that is driven by the hypoxia-inducible factor (HIF) pathway. The precise contribution of HIF-1a in the adaptive response, however, has not been determined. Here, we investigate how HIF influences hypoxic adaptation throughout Drosophila melanogaster development. We find that hypoxic-induced transcriptional changes are comprised of HIF-dependent and HIF-independent pathways that are distinct and separable. We show that normoxic set-points of carbohydrate metabolites are significantly altered in sima mutants and that these animals are unable to mobilize glycogen in hypoxia. …

Identification And Structural Characterization Of A New Three-Finger Toxin Hemachatoxin From Hemachatus Haemachatus Venom, Vallerinteavide Mavelli Girish, Sundramurthy Kumar, Lissa Joseph, Chacko Jobichen, R. Manjunatha Kini, J. Sivaraman

Biochemistry and Molecular Biology Publications

Snake venoms are rich sources of biologically active proteins and polypeptides. Three-finger toxins are non-enzymatic proteins present in elapid (cobras, kraits, mambas and sea snakes) and colubrid venoms. These proteins contain four conserved disulfide bonds in the core to maintain the three-finger folds. Although all three-finger toxins have similar fold, their biological activities are different. A new three-finger toxin (hemachatoxin) was isolated from Hemachatus haemachatus (Ringhals cobra) venom. Its amino acid sequence was elucidated, and crystal structure was determined at 2.43 Å resolution. The overall fold is similar to other three-finger toxins. The structure and sequence analysis revealed that the …