Robert D Goldman

Northwestern University, Evanston, Illinois, United States

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Publications (271)1935.73 Total impact

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    ABSTRACT: Giant Axonal Neuropathy (GAN) is a rare disease caused by mutations in the GAN gene which encodes gigaxonin, an E3 ligase adapter that targets intermediate filament (IF) proteins for degradation in numerous cell types including neurons and fibroblasts. The cellular hallmark of GAN pathology is the formation of large aggregates and bundles of IF. In this study we show that both the distribution and motility of mitochondria are altered in GAN fibroblasts and this is attributable to their association with vimentin IF aggregates and bundles. Transient expression of wild type gigaxonin in GAN fibroblasts reduces the number of IF aggregates and bundles, restoring mitochondrial motility. Conversely, silencing the expression of gigaxonin in control fibroblasts leads to changes in IF organization similar to GAN patients' fibroblasts and a coincident loss of mitochondrial motility. The inhibition of mitochondrial motility in GAN fibroblasts is not due to a global inhibition of organelle translocation, as lysosome motility is normal. Our findings demonstrate that it is the pathological changes in IF organization which cause the loss of mitochondrial motility.
    Preview · Article · Dec 2015 · Molecular biology of the cell
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    ABSTRACT: Macroautophagy (hereafter referred to as autophagy) is a catabolic membrane trafficking process that degrades a variety of cellular constituents and is associated with human diseases1, 2, 3. Although extensive studies have focused on autophagic turnover of cytoplasmic materials, little is known about the role of autophagy in degrading nuclear components. Here we report that the autophagy machinery mediates degradation of nuclear lamina components in mammals. The autophagy protein LC3/Atg8, which is involved in autophagy membrane trafficking and substrate delivery4, 5, 6, is present in the nucleus and directly interacts with the nuclear lamina protein lamin B1, and binds to lamin-associated domains on chromatin. This LC3–lamin B1 interaction does not downregulate lamin B1 during starvation, but mediates its degradation upon oncogenic insults, such as by activated RAS. Lamin B1 degradation is achieved by nucleus-to-cytoplasm transport that delivers lamin B1 to the lysosome. Inhibiting autophagy or the LC3–lamin B1 interaction prevents activated RAS-induced lamin B1 loss and attenuates oncogene-induced senescence in primary human cells. Our study suggests that this new function of autophagy acts as a guarding mechanism protecting cells from tumorigenesis.
    Full-text · Article · Nov 2015 · Nature
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    ABSTRACT: The nuclear lamina is a key structural element of the metazoan nucleus. However, the structural organization of the major proteins composing the lamina remains poorly defined. Using three-dimensional Structured Illumination Microscopy and computational image analysis, we have characterized the supramolecular structures of lamin A, C, B1 and B2 in mouse embryo fibroblast nuclei. Each isoform forms a distinct fiber meshwork, having comparable physical characteristics with respect to mesh edge length, mesh face area and shape, and edge connectivity to form faces. Some differences were found in face areas between isoforms due to variation in the edge lengths and number of edges per face, suggesting that each meshwork has somewhat unique assembly characteristics. In fibroblasts null for the expression of either lamins A/C or lamin B1, the remaining lamin meshworks are altered compared with the lamin meshworks in wild type nuclei or nuclei lacking lamin B2. Nuclei lacking LA/C exhibit slightly enlarged meshwork faces and some shape changes, whereas LB1-deficient nuclei exhibit primarily a substantial increase in face area. These studies demonstrate that individual lamin isoforms assemble into complex networks within the nuclear lamina and that A-type and B-type lamins have distinct roles in maintaining the organization of the nuclear lamina.
    Full-text · Article · Aug 2015 · Molecular biology of the cell
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    ABSTRACT: Intermediate Filaments (IFs) are composed of one or more members of a large family of cytoskeletal proteins, whose expression is cell and tissue type specific. Their importance in regulating the physiological properties of cells is becoming widely recognized in functions ranging from cell motility to signal transduction. IF proteins assemble into nanoscale biopolymers with unique strain hardening properties that are related to their roles in regulating the mechanical integrity of cells. Furthermore, mutations in the genes encoding IF proteins cause a wide range of human diseases. Due to the number of different types of IF proteins, we have limited this short review to cover structure and function topics mainly related to the simpler homopolymer IF networks comprised of vimentin, and specifically for diseases, the related muscle-specific desmin IF networks. Copyright © 2015, The American Society for Biochemistry and Molecular Biology.
    No preview · Article · May 2015 · Journal of Biological Chemistry
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    Sarah Köster · David A Weitz · Robert D Goldman · Ueli Aebi · Harald Herrmann
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    ABSTRACT: Intermediate filament proteins form filaments, fibers and networks both in the cytoplasm and the nucleus of metazoan cells. Their general structural building plan accommodates highly varying amino acid sequences to yield extended dimeric α-helical coiled coils of highly conserved design. These 'rod' particles are the basic building blocks of intrinsically flexible, filamentous structures that are able to resist high mechanical stresses, that is, bending and stretching to a considerable degree, both in vitro and in the cell. Biophysical and computer modeling studies are beginning to unfold detailed structural and mechanical insights into these major supramolecular assemblies of cell architecture, not only in the 'test tube' but also in the cellular and tissue context. Copyright © 2015 Elsevier Ltd. All rights reserved.
    Full-text · Article · Feb 2015 · Current Opinion in Cell Biology
  • Mikkel H. Jensen · Eliza J. Morris · Robert D. Goldman · David A. Weitz

    No preview · Article · Jan 2015 · Biophysical Journal
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    ABSTRACT: More than 20 mutations in the gene encoding A-type lamins (LMNA) cause progeria, a rare premature aging disorder. The major pathognomonic hallmarks of progeria cells are seen as nuclear deformations or blebs that are related to the redistribution of A- and B-type lamins within the nuclear lamina. However, the functional significance of these progeria-associated blebs remains unknown. We have carried out an analysis of the structural and functional consequences of progeria-associated nuclear blebs in dermal fibroblasts from a progeria patient carrying a rare point mutation p.S143F (C428T) in lamin A/C. These blebs form microdomains that are devoid of major structural components of the nuclear envelope (NE)/lamina including B-type lamins and nuclear pore complexes (NPCs) and are enriched in A-type lamins. Using laser capture microdissection and comparative genomic hybridization (CGH) analyses, we show that, while these domains are devoid of centromeric heterochromatin and gene-poor regions of chromosomes, they are enriched in gene-rich chromosomal regions. The active form of RNA polymerase II is also greatly enriched in blebs as well as nascent RNA but the nuclear co-activator SKIP is significantly reduced in blebs compared to other transcription factors. Our results suggest that the p.S143F progeria mutation has a severe impact not only on the structure of the lamina but also on the organization of interphase chromatin domains and transcription. These structural defects are likely to contribute to gene expression changes reported in progeria and other types of laminopathies.
    No preview · Article · Jan 2015 · Nucleus (Austin, Texas)
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    ABSTRACT: Nuclear lamins play important roles in the organization and structure of the nucleus; however, the specific mechanisms linking lamin structure to nuclear functions are poorly defined. We demonstrate that reducing nuclear lamin B1 expression by shRNA-mediated silencing in cancer cell lines to approximately 50% of normal levels causes a delay of the cell cycle and an accumulation of cells in early S phase. The S phase delay appears to be due to the stalling and collapse of replication forks. The double strand DNA breaks resulting from replication fork collapse were inefficiently repaired causing persistent DNA damage signaling and the assembly of extensive repair foci on chromatin. The expression of multiple factors involved in DNA replication and DNA repair by both non-homologous end joining and homologous repair is misregulated when lamin B1 levels are reduced. We further demonstrate that lamin B1 interacts directly with the promoters of some genes associated with DNA damage response and repair including BRCA1 and RAD51. Taken together the results suggest that the maintenance of lamin B1 levels is required for DNA replication and repair through regulating the expression of key factors involved in these essential nuclear functions. Copyright © 2014, American Society for Microbiology. All Rights Reserved.
    Full-text · Article · Dec 2014 · Molecular and Cellular Biology
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    ABSTRACT: Telomeres protect the ends of linear genomes, and the gradual loss of telomeres is associated with cellular ageing. Telomere protection involves the insertion of the 3' overhang facilitated by telomere repeat-binding factor 2 (TRF2) into telomeric DNA, forming t-loops. We present evidence suggesting that t-loops can also form at interstitial telomeric sequences in a TRF2-dependent manner, forming an interstitial t-loop (ITL). We demonstrate that TRF2 association with interstitial telomeric sequences is stabilized by co-localization with A-type lamins (lamin A/C). We also find that lamin A/C interacts with TRF2 and that reduction in levels of lamin A/C or mutations in LMNA that cause an autosomal dominant premature ageing disorder-Hutchinson Gilford Progeria Syndrome (HGPS)-lead to reduced ITL formation and telomere loss. We propose that cellular and organismal ageing are intertwined through the effects of the interaction between TRF2 and lamin A/C on chromosome structure.
    Full-text · Article · Nov 2014 · Nature Communications
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    ABSTRACT: This study demonstrates that the association of mitochondria with vimentin intermediate filaments (VIFs) measurably increases their membrane potential. This increase is detected by quantitatively comparing the fluorescence intensity of mitochondria stained with the membrane potential-sensitive dye tetramethylrhodamine-ethyl ester (TMRE) in murine vimentin-null fibroblasts with that in the same cells expressing human vimentin (∼35% rise). When vimentin expression is silenced by small hairpin RNA (shRNA) to reduce vimentin by 90%, the fluorescence intensity of mitochondria decreases by 20%. The increase in membrane potential is caused by specific interactions between a subdomain of the non-α-helical N terminus (residues 40 to 93) of vimentin and mitochondria. In rho 0 cells lacking mitochondrial DNA (mtDNA) and consequently missing several key proteins in the mitochondrial respiratory chain (ρ(0) cells), the membrane potential generated by an alternative anaerobic process is insensitive to the interactions between mitochondria and VIF. The results of our studies show that the close association between mitochondria and VIF is important both for determining their position in cells and their physiologic activity.-Chernoivanenko, I. S., Matveeva, E. A., Gelfand, V. I., Goldman, R. D., and Minin, A. A. Mitochondrial membrane potential is regulated by vimentin intermediate filaments. © FASEB.
    No preview · Article · Nov 2014 · The FASEB Journal
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    ABSTRACT: Molecular motors in cells typically produce highly directed motion; however, the aggregate, incoherent effect of all active processes also creates randomly fluctuating forces, which drive diffusive-like, nonthermal motion. Here, we introduce force-spectrum-microscopy (FSM) to directly quantify random forces within the cytoplasm of cells and thereby probe stochastic motor activity. This technique combines measurements of the random motion of probe particles with independent micromechanical measurements of the cytoplasm to quantify the spectrum of force fluctuations. Using FSM, we show that force fluctuations substantially enhance intracellular movement of small and large components. The fluctuations are three times larger in malignant cells than in their benign counterparts. We further demonstrate that vimentin acts globally to anchor organelles against randomly fluctuating forces in the cytoplasm, with no effect on their magnitude. Thus, FSM has broad applications for understanding the cytoplasm and its intracellular processes in relation to cell physiology in healthy and diseased states.
    No preview · Article · Aug 2014 · Cell
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    Mikkel H Jensen · Eliza J Morris · Robert D Goldman · David A Weitz
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    ABSTRACT: The semiflexible polymers filamentous actin (F-actin) and intermediate filaments (IF) both form complex networks within the cell, and together are key determinants of cellular stiffness. While the mechanics of F-actin networks together with stiff microtubules have been characterized, the interplay between F-actin and IF networks is largely unknown, necessitating the study of composite networks using mixtures of semiflexible biopolymers. We employ bulk rheology in a simplified in vitro system to uncover the fundamental mechanical interactions between networks of the 2 semiflexible polymers, F-actin and vimentin IF. Surprisingly, co-polymerization of actin and vimentin can produce composite networks either stronger or weaker than pure F-actin networks. We show that this effect occurs through steric constraints imposed by IF on F-actin during network formation and filament crosslinking, highlighting novel emergent behavior in composite semiflexible networks.
    Full-text · Article · May 2014 · Bioarchitecture
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    ABSTRACT: Nuclear lamins form the major structural elements comprising the nuclear lamina. While loss of nuclear structural integrity has been implicated as a key factor in the lamin A gene mutations causing laminopathies, the normal regulation of lamin A/C (LA/C) assembly and organization in interphase cells is still undefined. We assumed phosphorylation to be a major determinant, identifying 21 prime interphase phosphorylation sites, with 8 high turnover sites. The roles of these latter sites were examined by site-directed mutagenesis, followed by detailed microscopic analysis, including fluorescence recovery after photobleaching, fluorescence correlation spectroscopy, and nuclear extraction techniques. Results reveal three phosphorylation regions, each with dominant sites, together controlling LA/C structure and dynamics. Interestingly, two of these interphase sites are hyperphosphorylated in mitotic cells and one is within the sequence missing in progerin of the Hutchinson Gilford Progeria Syndrome. A model is presented where different phosphorylation combinations will yield markedly different effects on the assembly, subunit turnover, and mobility of LA/C between and within the lamina, the nucleoplasm, and the cytoplasm of interphase cells.
    Full-text · Article · Apr 2014 · Journal of Cell Science
  • Takeshi Shimi · Robert D Goldman
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    ABSTRACT: In mammalian cells, the nuclear lamina is composed of a complex fibrillar network associated with the inner membrane of the nuclear envelope. The lamina provides mechanical support for the nucleus and functions as the major determinant of its size and shape. At its innermost aspect it associates with peripheral components of chromatin and thereby contributes to the organization of interphase chromosomes. The A- and B-type lamins are the major structural components of the lamina, and numerous mutations in the A-type lamin gene have been shown to cause many types of human diseases collectively known as the laminopathies. These mutations have also been shown to cause a disruption in the normal interactions between the A and B lamin networks. The impact of these mutations on nuclear functions is related to the roles of lamins in regulating various essential processes including DNA synthesis and damage repair, transcription and the regulation of genes involved in the response to oxidative stress. The major cause of oxidative stress is the production of reactive oxygen species (ROS), which is critically important for cell proliferation and longevity. Moderate increases in ROS act to initiate signaling pathways involved in cell proliferation and differentiation, whereas excessive increases in ROS cause oxidative stress, which in turn induces cell death and/or senescence. In this review, we cover current findings about the role of lamins in regulating cell proliferation and longevity through oxidative stress responses and ROS signaling pathways. We also speculate on the involvement of lamins in tumor cell proliferation through the control of ROS metabolism.
    No preview · Article · Feb 2014 · Advances in Experimental Medicine and Biology
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    Yuxuan Guo · Youngjo Kim · Takeshi Shimi · Robert D Goldman · Yixian Zheng
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    ABSTRACT: The nuclear lamina (NL) consists of lamin polymers and proteins that bind to the polymers. Disruption of NL proteins such as lamin and emerin leads to developmental defects and human diseases. However, the expression of multiple lamins, including lamin-A/C, lamin-B1, and lamin-B2, in mammals has made it difficult to study the assembly and function of the NL. Consequently, it remains unclear whether different lamins depend on one another for proper NL assembly and which NL functions are shared by all lamins or are specific to one lamin. Using mouse cells deleted of all or different combinations of lamins, we demonstrate that the assembly of each lamin into NL depends primarily on the lamin concentration present in the nucleus. When expressed at sufficiently high levels, each lamin alone can assemble into an evenly organized NL, which is in turn sufficient to ensure the even distribution of the nuclear pore complexes (NPC). By contrast, only lamin-A can ensure the localization of emerin within the NL. Thus, when investigating the role of the NL in development and disease, it is critical to determine the protein levels of relevant lamins and the intricate shared or specific lamin functions in the tissue of interest.
    Preview · Article · Feb 2014 · Molecular biology of the cell
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    Full-text · Dataset · Dec 2013
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    ABSTRACT: Significant efforts have addressed the role of vimentin intermediate filaments (VIF) in cell motility, shape, adhesion and their connections to microfilaments (MF) and microtubules (MT). The present work uses micropatterned substrates to control the shapes of mouse fibroblasts and demonstrates that the cytoskeletal elements are dependent on each other and that unlike MF, VIF are globally controlled. For example, both square and circle shaped cells have a similar VIF distribution while MF distributions in these two shapes are quite different and depend on the curvature of the shape. Furthermore, in asymmetric and polarized shaped cells VIF avoid the sharp edges where MF are highly localized. Experiments with vimentin null mouse embryonic fibroblasts (MEFs) adherent to polarized (teardrop) and un-polarized (dumbbell) patterns show that the absence of VIF alters microtubule organization and perturbs cell polarity. The results of this study also demonstrate the utility of patterned substrates for quantitative studies of cytoskeleton organization in adherent cells.
    Full-text · Article · Nov 2013 · Biomaterials
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    ABSTRACT: The mechanical properties of a cell determine many aspects of its behavior, and these mechanics are largely determined by the cytoskeleton. Although the contribution of actin filaments and microtubules to the mechanics of cells has been investigated in great detail, relatively little is known about the contribution of the third major cytoskeletal component, intermediate filaments (IFs). To determine the role of vimentin IF (VIF) in modulating intracellular and cortical mechanics, we carried out studies using mouse embryonic fibroblasts (mEFs) derived from wild-type or vimentin(-/-) mice. The VIFs contribute little to cortical stiffness but are critical for regulating intracellular mechanics. Active microrheology measurements using optical tweezers in living cells reveal that the presence of VIFs doubles the value of the cytoplasmic shear modulus to ∼10 Pa. The higher levels of cytoplasmic stiffness appear to stabilize organelles in the cell, as measured by tracking endogenous vesicle movement. These studies show that VIFs both increase the mechanical integrity of cells and localize intracellular components.
    Full-text · Article · Oct 2013 · Biophysical Journal
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    Veronika Butin-Israeli · Stephen A Adam · Robert D Goldman
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    ABSTRACT: The nuclear lamins play important roles in the structural organization and function of the metazoan cell nucleus. Recent studies on B-type lamins identified a requirement for lamin B1 (LB1) in the regulation of cell proliferation in normal diploid cells. In order to further investigate the function of LB1 in proliferation, we disrupted its normal expression in U-2 OS human osteosarcoma and other tumor cell lines. Silencing LB1 expression induced G1 cell cycle arrest without significant apoptosis. The arrested cells are unable to mount a timely and effective response to DNA damage induced by UV irradiation. Several proteins involved in the detection and repair of UV damage by the nucleotide excision repair (NER) pathway are down-regulated in LB1 silenced cells including DDB1, CSB and PCNA. We propose that LB1 regulates the DNA damage response to UV irradiation by modulating the expression of specific genes and activating persistent DNA damage signaling. Our findings are relevant to understanding the relationship between the loss of LB1 expression, DNA damage signaling, and replicative senescence.
    Full-text · Article · Jul 2013 · PLoS ONE
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    Puneet Opal · Robert D Goldman
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    ABSTRACT: Giant axonal neuropathy (GAN)11. Mahammad S, Murthy SN, Didonna A, Grin B, Israeli E, Perrot R, et al. Giant axonal neuropathy-associated gigaxonin mutations impair intermediate filament protein degradation. J Clin Invest 2013; 123:1964 - 75;; PMID: 23585478 [CrossRef]View all references is a rare autosomal recessive neurological disorder caused by mutations in the GAN gene that encodes gigaxonin, a member of the BTB/Kelch family of E3 ligase adaptor proteins.11. Mahammad S, Murthy SN, Didonna A, Grin B, Israeli E, Perrot R, et al. Giant axonal neuropathy-associated gigaxonin mutations impair intermediate filament protein degradation. J Clin Invest 2013; 123:1964 - 75;; PMID: 23585478 [CrossRef]View all references This disease is characterized by the aggregation of Intermediate Filaments (IF)—cytoskeletal elements that play important roles in cell physiology including the regulation of cell shape, motility, mechanics and intra-cellular signaling. Although a range of cell types are affected in GAN, neurons display the most severe pathology, with neuronal intermediate filament accumulation and aggregation; this in turn causes axonal swellings or “giant axons.” A mechanistic understanding of GAN IF pathology has eluded researchers for many years. In a recent study11. Mahammad S, Murthy SN, Didonna A, Grin B, Israeli E, Perrot R, et al. Giant axonal neuropathy-associated gigaxonin mutations impair intermediate filament protein degradation. J Clin Invest 2013; 123:1964 - 75;; PMID: 23585478 [CrossRef]View all references we demonstrate that the normal function of gigaxonin is to regulate the degradation of IF proteins via the proteasome. Our findings present the first direct link between GAN mutations and IF pathology; moreover, given the importance of IF aggregations in a wide range of disease conditions, our findings could have wider ramifications.
    Preview · Article · Jun 2013

Publication Stats

20k Citations
1,935.73 Total Impact Points


  • 1984-2015
    • Northwestern University
      • • Department of Cell and Molecular Biology
      • • Feinberg School of Medicine
      Evanston, Illinois, United States
    • University of Colorado at Boulder
      Boulder, Colorado, United States
    • National Institutes of Health
      • Branch of Dermatology
      베서스다, Maryland, United States
  • 2004
    • French National Centre for Scientific Research
      Lutetia Parisorum, Île-de-France, France
    • University of Turku
      • Department of Biology
      Turku, Western Finland, Finland
  • 1983-2003
    • University of Illinois at Chicago
      • Department of Anatomy and Cell Biology (Chicago)
      Chicago, Illinois, United States
  • 2002
    • Hebrew University of Jerusalem
      Yerushalayim, Jerusalem, Israel
  • 1999
    • University of Pennsylvania
      • Institute for Medicine and Engineering
      Philadelphia, PA, United States
  • 1998
    • Columbia University
      • Department of Pathology & Cell Biology
      New York, New York, United States
    • Johns Hopkins University
      • Department of Biological Chemistry
      Baltimore, Maryland, United States
    • University of California, Davis
      • Department of Mechanical and Aerospace Engineering
      Davis, California, United States
    • Howard Hughes Medical Institute
      Ashburn, Virginia, United States
  • 1997
    • Georg-August-Universität Göttingen
      Göttingen, Lower Saxony, Germany
  • 1992
    • University of Rochester
      Rochester, New York, United States
  • 1991
    • University of Cincinnati
      Cincinnati, Ohio, United States
  • 1989
    • University of Wisconsin–Madison
      • Laboratory of Cell and Molecular Biology
      Madison, Wisconsin, United States
    • University of Virginia
      • Department of Biology
      Charlottesville, Virginia, United States
  • 1985
    • Marine Biological Laboratory
      Falmouth, Massachusetts, United States
    • The University of Chicago Medical Center
      • Department of Cell Biology and Anatomy
      Chicago, Illinois, United States
  • 1974-1982
    • Carnegie Mellon University
      • Department of Biological Sciences
      Pittsburgh, Pennsylvania, United States
  • 1981
    • National Cancer Institute (USA)
      • Dermatology Branch
      베서스다, Maryland, United States
  • 1970-1979
    • Case Western Reserve University
      • Department of Biology
      Cleveland, Ohio, United States
    • Institute of Human Virology
      Maryland City, Maryland, United States
  • 1974-1976
    • Cold Spring Harbor Laboratory
      Cold Spring Harbor, New York, United States