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THE 3' UNTRANSLATED REGIONS OF 2 RELATED MESSENGER-RNAS CONTAIN AN ELEMENT HIGHLY REPEATED IN THE SEA-URCHIN GENOME

NUCLEIC ACIDS RESEARCH 1982年第23期10卷 7829-7842页

作者： CARPENTER, CD BRUSKIN, AM SPAIN, LM ELDON, ED KLEIN, WH Program in Molecular Cellular and Developmental Biology Department of Biology Indiana UniversityBloomington IN 47405 USA

Two closely related cDNA clones, pSpec1 and pSpec2, specifying two developmentally regulated tissue specific mRNAs from sea urchin embryos were used to probe a sea urchin genomic lambda library. Screening 10,000 phage by plaque hybridization yielded several hundred positive signals. With more stringent wash procedures, only two to three phage were positive. Three of these phage, one isolated by stringent wash procedures and two isolated by standard wash procedures were further investigated by restriction analysis, RNA gel blots, and DNA sequencing. The phage isolated by the stringent wash procedure appears to be a gene coding for the Specl mRNA. The other phage contain only partial homology to pSpec1 and pSpec2, 150 to 200 base pairs of the 3' untranslated region of the Spec1 and Spec2 mRNAs. It is concluded that the Spec1 and Spec2 mRNAs contain a highly repetitive element near their 3' end. The element is present at 2000 to 3000 copies per genome and may be transcribed at some sites other than those coding for the Spec1 and Spec2 genes. The possible function and evolutionary origin of the repetitive element is discussed.

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FOLLICLE CELL CALMODULIN IN BLATTELLA-GERMANICA TRANSCRIPT ACCUMULATION DURING VITELLOGENESIS IS REGULATED BY JUVENILE-HORMONE

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DEVELOPMENTAL biology 1995年第2期170卷 314-320页

作者： IYENGAR, AR KUNKEL, JG Molecular and Cellular Biology Program University of Massachusetts Amherst Massachusetts 01003

There is abundant calmodulin (CaM) in the oocytes and eggs of B, germanica. Whether oocytes accumulate CaM for immediate use or use at a later stage in their development is still unknown. We show that isolated follicle cells accumulate more CaM transcripts per unit RNA than any other control tissue. CaM transcript increases exponentially 4800-fold in follicles during the 96-hr vitellogenic period in the absence of cell division. This includes a 32-fold increase in total follicle RNA during the period and an 150-fold increase in relative titer of the CaM transcript. In comparison, levels of actin transcripts increase exponentially 1200-fold during the same developmental period. On the other hand fat body tissue shows little relative increase of CaM transcripts despite a 4-fold increase in total RNA over the 4-day developmental period. Both the CaM and actin transcripts are more highly concentrated in the Bay 4 follicle cell layer, being found in 84- and 33-fold greater titer, respectively, than in fat body RNA. Deprivation of juvenile hormone (JH), by head ligation, not only causes atresia of the follicles, but also reduces accumulated CaM transcripts. Reestablishing JH titer by injection allows a selected population of follicles to develop to full size and also reinstates CaM transcript levels above that of unligated controls within 24 hr. (C) 1995 Academic Press, Inc.

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EFFECTS OF SUBSTANCE-P ON SECRETION OF CATECHOLAMINES FROM POPULATIONS OF BOVINE CHROMAFFIN CELLS AND ON CALCIUM TRANSIENTS IN INDIVIDUAL CELLS

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ANNALS OF THE NEW YORK ACADEMY OF SCIENCES 1991年第1期632卷 241-248页

作者： KENTBRAUN, JA LYFORD, LK GROSS, DJ WESTHEAD, EW Program in Molecular and Cellular Biology University of Massachusetts Amherst 01003.

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Mice with lymphatic dysfunction develop pathogenic lung tertiary lymphoid organs that model an autoimmune emphysema phenotype of COPD

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American Journal of Physiology - Lung cellular and molecular Physiology 2025年第1期328卷 L1-L14页

作者： Summers, Barbara Kim, Kihwan Trivedi, Anjali Lu, Tyler M. Houghton, Sean Palmer-Johnson, Jade Rojas-Quintero, Joselyn Cala-Garcia, Juan Pannellini, Tania Polverino, Francesca Lis, Raphaël Reed, Hasina Outtz Division of Pulmonary and Critical Care Department of Medicine Weill Cornell Medicine New York NY United States Division of Regenerative Medicine Department of Medicine Ansary Stem Cell Institute Weill Cornell Medicine New York NY United States Ronald O. Perelman and Claudia Cohen Center for Reproductive Medicine Weill Cornell Medicine New York NY United States Molecular and Cellular Biology Program SUNY Downstate School of Graduate Studies Brooklyn NY United States Department of Medicine Section of Pulmonary Critical Care and Sleep Medicine Baylor College of Medicine Houston TX United States Department of Pathology Weill Cornell Medicine New York NY United States Department of Cell and Developmental Biology Weill Cornell Medicine New York NY United States

We have previously shown that mice with a loss of C-type lectin-like type II (CLEC2), which have lymphatic dysfunction due to the role of CLEC2 in platelets for maintaining separation between the venous and lymphatic system, develop lung tertiary lymphoid organ (TLO) formation and lung injury that resembles an emphysema phenotype of chronic obstructive pulmonary disease (COPD). We now sought to investigate whether and how TLOs in these mice may play a pathogenic role in lung injury that is relevant to human disease. We found that inhibiting TLO formation using an anti-CD20 antibody in CLEC2-deficient mice partially blocked the development of emphysema. TLOs in CLEC2-deficient mice were rich in plasma cells and were a source of a broad array of autoantibodies. Chronic cigarette smoke exposure increased the size and number of lung TLOs in CLEC2-deficient mice and was associated with increased markers of antigen presentation and maturation, leading to increased autoantibody deposition. Using lung tissue from patients with COPD, we found an increase in lymphatic markers in patients with an emphysema phenotype and autoreactive TLOs compared with patients with COPD without emphysema that lack prominent TLOs. Taken together, these results demonstrate that emphysema in mice with lymphatic dysfunction can be partially rescued by blocking TLO formation and that these TLOs are the source of autoantibodies that are exacerbated by cigarette smoke. Our work suggests that lymphatic dysfunction in mice may recapitulate some aspects of an autoimmune emphysema phenotype that is seen in a subset of patients with COPD. Copyright © 2025 The Authors.

关键词： autoimmunity COPD emphysema lymphatic vasculature tertiary lymphoid organs

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DELAYED RECRUITMENT OF MATERNAL HISTONE H-3 MESSENGER-RNA IN SEA-URCHIN EMBRYOS

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NATURE 1981年第5822期292卷 477-478页

作者： WELLS, DE SHOWMAN, RM KLEIN, WH RAFF, RA Program in Molecular Cellular and Developmental Biology Department of Biology Indiana University Bloomington USA

During the early stages of embryogenesis, the sea urchin embryo uses maternally synthesized mRNA stored in the egg as inactive messenger ribonucleoproteins (mRNPs)1. Release of this mRNA for translation allows the embryo to develop even in the absence of new mRNA synthesis. Comparison using two-dimensional gel electrophoresis shows that the same spectrum of prevalent proteins are synthesized by eggs and zygotes 30–60 min after fertilization2. Thus, unlike the case of the clamSpisulain which there are several prominent changes in translation associated with fertilization3, sea urchins have been thought to show little or no regulation of translation of specific mRNA sequences. Histone mRNAs are major components of the mRNA pool, comprising as much as 4–8% of the total mRNA of eggs4, yet their products, histones, would not normally be detectable on two-dimensional gels. We report here that mRNA complementary to a histone H3 cloned probe remains in the inactive mRNA pool for 90 min after fertilization before it begins to be translated. This is long after the rapid increase in overall protein synthesis, using stored mRNAs, has begun. As we can demonstrate no significant synthesis of histone H3 mRNA during this period, stored histone H3 mRNA must be subject to sequence-specific translational regulation.

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molecular DETAILS OF TOMATO EXTENSIN AND GLYCINE-RICH PROTEIN GENE-EXPRESSION

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PLANT molecular biology 1992年第2期19卷 205-215页

作者： SHOWALTER, AM BUTT, AD KIM, S Department of Botany Molecular and Cellular Biology Program Ohio University Athens USA

In a recent publication (Plant molecular biology 16: 547-565 (1991)), Showalter et al. described the isolation and initial characterization of fifteen extensin and extensin-like tomato cDNAs. These cDNAs were determined to fall into five distinct classes;class I and II clones encoded extensins, class III and V clones encoded glycine-rich proteins (GRPs), and class IV clones encoded a portion of a GRP sequence on one DNA strand and a portion of an extensin sequence on the other DNA strand. In this publication, a more detailed analysis of the expression of these cDNA classes was performed with respect to wounding in various tomato organs, development, kinetics and systemic extent of the wound response, ethylene treatment, abscisic acid (ABA) treatment, and drought stress by using RNA gel blot hybridizations. In general, extension gene expression was readily detected in stems and roots, but not in leaves. With both class I and II extensin cDNA probes, wound-induced accumulation of mRNA in stems was first detected between 4 and 8 h after wounding with maximal accumulation occurring after 12 h. Moreover, these extensin wound responses were detected locally at the wound site but not systemically. Expression of the class III GRP was largely limited to wounded stem tissue. Initial detection and maximal accumulation of the class III GRP mRNA was similar to the extensins mRNAs, however, this GRP wound response occurred both locally and systemically. Additionally, abscisic acid treatment and drought stress resulted in the marked accumulation the class III GRP mRNA in tomato stems, but did not alter the expression of the other cDNA classes. In contrast, expression of the class V GRP occurred in stems and roots and to a lesser extent in leaves and decreased in response to wounding over a 24 h time period. The class V GRP wound response was further characterized by an early, transient accumulation of mRNA occurring 2-4 h after wounding in stems and by its local nature.

关键词： EXTENSIN GLYCINE-RICH PROTEIN HYDROXYPROLINE-RICH GLYCOPROTEIN TOMATO WATER STRESS WOUNDING

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IDENTIFICATION AND GENETIC LOCALIZATION OF MESSENGER-RNA FROM OVARIAN FOLLICLE CELLS OF DROSOPHILA-MELANOGASTER

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CELL 1979年第3期16卷 589-598页

作者： SPRADLING, AC MAHOWALD, AP Program in Molecular Cellular and Developmental Biology Department of Biology Indiana University Bloomington Indiana 47401 USA

RNA synthesis in ovarian follicles of D. melanogaster was studied by methods which eliminate experimentally induced alterations in gene expression. Gel electrophoresis of follicular RNA, labeled after injection of precursors into females, revealed qualitative and quantitative differences in synthesis during the course of oogenesis. A highly heterogeneous group of poly(A)-containing RNA is produced during much of the course of follicular development. Post-vitellogenic stages synthesize a small number of stage-specific poly(A)-containing RNA. During this period, RNA synthesis takes place primarily in the follicle cells, which are engaged in the production of the endochorion and exochorion. Two intense bands of nonmitochondrial poly(A)+ RNA are labeled between stage 11 and early stage 13. The synthesis of a more heterogeneous group of very small poly(A)-containing RNA characterizes the last part of oogenesis, stages 13 and 14. Evidence is presented to show that these RNA are specifically localized in the follicle cells of the egg chamber. They apparently represent mRNA for chorion proteins. In situ hybridization of preparations of late stage poly(A)-containing RNA to salivary gland chromosomes revealed 2 major sites of complementarity, 7E11 and 12E, and several minor sites. Experiments om which RNA were separated on gels prior to hybridization in situ suggested that both the major stage 12-specific RNA bands contained molecules which were complementary to DNA in the 7E11 region. This site is within a small chromosomal interval known to contain the gene ocelliless. Females homozygous for ocelliless produced structurally abnormal chorions (Johnson and King, 1974).

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ACCUMULATION IN EMBRYOGENESIS OF 5 MESSENGER-RNAS ENRICHED IN THE ECTODERM OF THE SEA-URCHIN PLUTEUS

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DEVELOPMENTAL biology 1981年第2期87卷 308-318页

作者： BRUSKIN, AM TYNER, AL WELLS, DE SHOWMAN, RM KLEIN, WH Program in Molecular Cellular and Developmental Biology Department of Biology Indiana University Bloomington Indiana 47401 USA

Five cDNA clones representing mRNA enriched in the ectoderm of the sea urchin pluteus were isolated from a library of 100 pluteus cDNA clones. RNA gel blots were used to show that the RNA are present at levels 3-20 times higher in the ectoderm than in the endoderm/mesoderm. Based on genomic DNA gel blots the 5 different mRNA were placed into 4 groups. Of the mRNA 3 belong to separate groups, and the remaining 2 mRNA are members of a 4th group. The accumulation of the mRNA during embryogenesis was measured using total cellular RNA from unfertilized eggs, 16-cell, mesenchyme blastula, mid-gastrula and pluteus-stage embryos. All mRNA are present at very low or undetectable levels in the unfertilized egg and 16-cell embryo. The amounts then increase sharply and reach a maximum at blastula or gastrula depending on the particular RNA, and then decline significantly by pluteus. In most RNA levels change > 100-fold in < 48 h of development. This accumulation is due to transcription during embryogenesis. The mRNA are found on polyribosomes at blastula and pluteus stages, but the nature of the proteins they encode is unknown.

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Identification and characterization of GIP1, an Arabidopsis thaliana protein that enhances the DNA binding affinity and reduces the oligomeric state of G-box binding factors

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Cell Research 2005年第8期15卷 567-575页

作者： Paul C. SEHNKE,Beth J. LAUGHNER,Carla R. LYERLY LINEBARGER,William B. GURLEY,Robert J. FERL Program in Plant Cellular and Molecular Biology Department of Horticultural Sciences University of Florida Gainesville FL USA 32611 Program in Plant Cellular and Molecular Biology Department of Horticultural Sciences University of Florida Gainesville FL USA 32611 Program in Plant Cellular and Molecular Biology Department of Horticultural Sciences University of Florida Gainesville FL USA 32611 Department of Microbiology and Cell Science University of Florida Gainesville FL USA 32611 Program in Plant Cellular and Molecular Biology Department of Horticultural Sciences University of Florida Gainesville FL USA 32611

Environmental control of the alcohol dehydrogenase (Adh) and other stress response genes in plants is in part brought about by transcriptional regulation involving the G-box cis-acting DNA element and bZIP G-box Binding Factors (GBFs). The mechanisms of GBF regulation and requirements for additional factors in this control process are not well understood. In an effort to identify potential GBF binding and control partners, maize GBF1 was used as bait in a yeast two-hybrid screen of an A. thaliana cDNA library. GBF Interacting Protein 1 (GIP1) arose from the screen as a 496 amino acid protein with a predicted molecular weight of 53,748 kDa that strongly interacts with GBFs. Northern analysis of A. thaliana tissue suggests a 1.8-1.9 kb GIP1 transcript, predominantly in roots. Immunolocalization studies indicate that GIP1 protein is mainly localized to the nucleus. In vitro electrophoretic mobility shift assays using an Adh G-box DNA probe and recombinant A. thaliana GBF3 or maize GBF1, showed that the presence of GIP1 resulted in a tenfold increase in GBF DNA binding activity without altering the migration, suggesting a transient association between GIP1 and GBF. Addition of GIP1 to intentionally aggregated GBF converted GBF to lower molecular weight macromolecular complexes and GIP1 also refolded denatured rhodanese in the absence of ATP. These data suggest GIP1 functions to enhance GBF DNA binding activity by acting as a potent nuclear chaperone or crowbar, and potentially regulates the multimeric state of GBFs, thereby contributing to bZIP-mediated gene regulation.

关键词： G-box binding factor interacting protein nuclear chaperone DNA binding affinity.

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Viroids:Small Probes for Exploring the Vast Universe of RNA Trafficking in Plants

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Journal of Integrative Plant biology 2010年第1期52卷 28-39页

作者： Ying Wang Biao Ding Department of Plant Cellular and Molecular Biology and Plant Biotechnology Center The Ohio State University The Center for RNA Biology The Ohio State University Molecular Cellular and DeVelopmental Biology Program The Ohio State University Columbus OH 43210 USA

Cell-to-cell and long-distance trafficking of RNA is a rapidly evolving frontier of integrative plant biology that broadly impacts studies on plant growth and development, spread of infectious agents and plant defense responses. The fundamental questions being pursued at the forefronts revolve around function, mechanism and evolution. In the present review, we will first use specific examples to illustrate the biological importance of cell-to-cell and long-distance trafficking of RNA. We then focus our discussion on research findings obtained using viroids that have advanced our understanding of the underlying mechanisms involved in RNA trafficking. We further use viroid examples to illustrate the great diversity of trafficking machinery evolved by plants, as well as the promise for new insights in the years ahead. Finally, we discuss the prospect of integrating findings from different experimental systems to achieve a systems-based understanding of RNA trafficking function, mechanism and evolution.

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