Kathryn G Miller

Kathryn G Miller
Washington University in St. Louis | WUSTL , Wash U · Department of Biology

Ph.D.

About

60
Publications
6,871
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3,572
Citations
Citations since 2017
2 Research Items
358 Citations
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2017201820192020202120222023020406080
2017201820192020202120222023020406080
2017201820192020202120222023020406080
Introduction
Actin structures play important roles in differentiation and in specialized cell functions. how they form in the correct place, proper time and with proper organization for their roles in many different contexts is not very well understood. We use Drosophila as a model organism to study the mechanisms that control actin structure assembly, organization and function in different types of cells.
Additional affiliations
January 1989 - present
January 1989 - present
Washington University in St. Louis
Position
  • Professor
January 1981 - December 1989
University of California, San Francisco
Education
September 1974 - December 1980
Johns Hopkins Medicine
Field of study
  • Biochemistry

Publications

Publications (60)
Article
Full-text available
A useful model for determining the mechanisms by which actin and actin binding proteins control cellular architecture is the Drosophila melanogaster process of spermatogenesis. During the final step of spermatogenesis, 64 syncytial spermatids individualized as stable actin cones move synchronously down the axonemes and remodel the membranes. To ide...
Article
Full-text available
Myosin VI (MVI) is a versatile actin-based motor protein that has been implicated in a variety of different cellular processes, including endo- and exocytic vesicle trafficking, Golgi morphology, and actin structure stabilization. A role for MVI in crucial actin-based processes involved in sperm maturation was demonstrated in Drosophila. Because of...
Article
Full-text available
The PULSE Vision & Change Rubrics,version 1.0,assess life sciences departments’ progress toward implementation of the principles of the Vision and Change report. This paper reports on the development of the rubrics,their validation,and their reliability in measuring departmental change aligned with the Vision and Change recommendations. The rubrics...
Article
Collaboration among faculty and teaching center staff has produced in-class, active-learning methods that help students learn visualization, problem-solving, critical thinking, and communication skills as they develop disciplinary knowledge. These methods include instructor and student use of technology tools—or tools that are unobtrusive in the cl...
Data
Ultrastructure of actin cones is altered when myosin VI is mislocalized in wild-type animals. Examples of S1 decorated actin cones when GFP-HNPMD (Gtail deleted) transgene is expressed in a wild-type animal. Big arrow indicates the direction of cone movement and small arrows indicate asymmetry of cone front domains. mi, mitochondria; ax, axoneme. B...
Data
Ultrastructure of actin cones is altered when myosin VI is mislocalized in mutant animals. Examples of S1 decorated cones when GFP-HNPMD (Gtail deleted) transgene is expressed in myosin VI mutant background. (insets, i–iii), Small regions of actin cones at high magnification. Big arrow indicates the direction of cone movement and small arrows indic...
Article
Full-text available
Actin structures are often stable, remaining unchanged in organization for the lifetime of a differentiated cell. Little is known about stable actin structure formation, organization, or maintenance. During Drosophila spermatid individualization, long-lived actin cones mediate cellular remodeling. Myosin VI is necessary for building the dense meshw...
Article
Stable actin structures play important roles in the development and specialization of differentiated cells. How these structures form, are organized, and are used to mediate physiological processes is not well understood in most cases. In Drosophila testis, stable actin structures, called actin cones, mediate spermatid individualization, a large-sc...
Article
Full-text available
Biology education research (BER) can be a major contributor to the herculean task of modernizing and transforming biology education. However, as researchers in a relatively young and still small field, BER practitioners now find themselves fragmented across 64 biology-related societies and lacking agreement on a core research agenda, a convenient p...
Article
Full-text available
Myosin VI is a pointed-end-directed actin motor that is thought to function as both a transporter of cargoes and an anchor, capable of binding cellular components to actin for long periods. Dimerization via a predicted coiled coil was hypothesized to regulate activity and motor properties. However, the importance of the coiled-coil sequence has not...
Article
Full-text available
During spermatid individualization in Drosophila, actin structures (cones) mediate cellular remodeling that separates the syncytial spermatids into individual cells. These actin cones are composed of two structural domains, a front meshwork and a rear region of parallel bundles. We show here that the two domains form separately in time, are regulat...
Article
Full-text available
Myosin VI is an actin-based motor that has been implicated in many cellular processes. Studies in vertebrates have demonstrated that animals lacking this ubiquitously expressed myosin are viable. However in Drosophila, myosin VI loss of function has been thought to be lethal. We show here that complete loss of myosin VI is not lethal in flies and t...
Article
Full-text available
Drosophila melanogaster bristle development is dependent on actin assembly, and prominent actin bundles form against the elongating cell membrane, giving the adult bristle its characteristic grooved pattern. Previous work has demonstrated that several actin-regulating proteins are required to generate normal actin bundles. Here we have addressed ho...
Article
Full-text available
Myosin VI, a ubiquitously expressed unconventional myosin, has roles in a broad array of biological processes. Unusual for this motor family, myosin VI moves toward the minus (pointed) end of actin filaments. Myosin VI has two light chain binding sites that can both bind calmodulin (CaM). However unconventional myosins could use tissue-specific lig...
Article
Full-text available
Here, we demonstrate a new function of myosin VI using observations of Drosophila spermatid individualization in vivo. We find that myosin VI stabilizes a branched actin network in actin structures (cones) that mediate the separation of the syncytial spermatids. In a myosin VI mutant, the cones do not accumulate F-actin during cone movement, wherea...
Article
Full-text available
Myosin VI is a member of a superfamily of actin-based motors with at least 18 different sub-types or classes. Myosins are best known as proteins that use ATP-hydrolysis-mediated conformational changes to move along actin filaments. Because of this property, some myosins, including myosins I, V, and VI, are thought to be transporters of vesicle or p...
Article
Full-text available
Myosin VI can move along actin filaments to serve as a transport motor. It is also thought to anchor vesicles or proteins to actin. How these two diverse activities, which require very different modes of interaction with actin, are mediated is not understood. Using single molecule observations, Altman et al. (2004 [this issue of Cell]) demonstrate...
Article
Full-text available
In order to better understand the mechanism of sperm individualization during spermatogenesis in Drosophila melanogaster, we have developed an in vitro culture system in which we can perform live observation of individualization in isolated cysts. The whole process of individualization, during which a bundle of 64 syncytial spermatids is separated...
Article
Three groups have recently characterized defects arising in development owing to mutations in the gene encoding Dmoesin, which is the sole ezrin-radixin-moesin (ERM) protein in Drosophila. Previously, studies in cultured mammalian cells suggested that ERM proteins are important for actin-membrane associations. However, mutations in moesin and radix...
Article
Full-text available
Profilin is a well-characterized protein known to be important for regulating actin filament assembly. Relatively few studies have addressed how profilin interacts with other actin-binding proteins in vivo to regulate assembly of complex actin structures. To investigate the function of profilin in the context of a differentiating cell, we have stud...
Article
Full-text available
Myosin VI has been implicated in membrane dynamics in several organisms. The mechanism of its participation in membrane events is not clear. We have used spermatogenesis in Drosophila to investigate myosin VI's in vivo role. We demonstrate that myosin VI colocalizes with and is required for the accumulation of the actin polymerization regulatory pr...
Article
Full-text available
Actin is a highly conserved protein found in all eukaryotic organisms. Most organisms have multiple cytoplasmic actin genes that encode isoforms with slightly different amino acid sequences. These different isoforms are coexpressed in many cell types. Why organisms have multiple very similar cytoplasmic actin genes is unclear. We have addressed thi...
Article
Natural populations of sexually reproducing Drosophila mercatorum are capable of a very low rate of parthenogenesis, but this mode of reproduction has apparently never characterized an entirely asexual population in this species. The high abortion rate observed in laboratory parthenogenetic lines suggests that developmental constraints may cause th...
Article
Full-text available
Developmental instability is particularly pronounced in parthenogenetic strains of Drosophila mercatorum. All parthenogenetically produced eggs in a given strain have the same genotype, but even when reared in the same environment, only approximately 5% of the eggs initiating development ever reach adulthood. A sexual analogue of a parthenogenetic...
Article
Full-text available
Two studies characterizing Drosophila Arp2/3 complex and Scar mutants demonstrate that assembly of some actin structures in nonmotile cells of multicellular organisms utilizes the same proteins as are important for actin assembly in motile cells. These studies also show that assembly of other actin structures is independent of these proteins, sugge...
Article
Full-text available
We have identified partial loss of function mutations in class VI unconventional myosin, 95F myosin, which results in male sterility. During spermatogenesis the germ line precursor cells undergo mitosis and meiosis to form a bundle of 64 spermatids. The spermatids remain interconnected by cytoplasmic bridges until individualization. The process of...
Article
Localization of mRNAs is one of many aspects of cellular organization that requires the cytoskeleton. In Drosophila, microtubules are known to be required for correct localization of developmentally important mRNAs and proteins during oogenesis; however, the role of the actin cytoskeleton in localization is less clear. Furthermore, it is not known...
Article
Full-text available
Coordination of cellular organization requires the interaction of the cytoskeletal filament systems. Recently, several lines of investigation have suggested that transport of cellular components along both microtubules and actin filaments is important for cellular organization and function. We report here on molecules that may mediate coordination...
Article
Regulation of actin filament length and orientation is important in many actin-based cellular processes. This regulation is postulated to occur through the action of actin-binding proteins. Many actin-binding proteins that modify actin in vitro have been identified, but in many cases, it is not known if this activity is physiologically relevant. Ca...
Article
Full-text available
The 95F myosin, a class VI unconventional myosin, associates with particles in the cytoplasm of the Drosophila syncytial blastoderm and is required for the ATP- and F-actin-dependent translocation of these particles. The particles undergo a cell cycle-dependent redistribution from domains that surround each nucleus in interphase to transient membra...
Article
Myosins are actin-activated ATPases that are able to translocate along actin filaments using energy derived from ATP hydrolysis. Non-muscle cells contain conventional myosins, which are similar in sequence and structure to muscle myosin, and a number of unconventional myosins whose head sequences are similar but tail sequences are unrelated to conv...
Article
In this summary of methods, we have attempted to present a series of protocols relevant to the study of proteins that associate with each other inside cells. These techniques have found their main application, in our hands, to cytoskeletal structures, but the utility of these techniques is limited to this application. As we learn more about the phy...
Article
Full-text available
In the syncytial blastoderm stage of Drosophila embryogenesis, dome-shaped actin "caps" are observed above the interphase nuclei. During mitosis, this actin rearranges to participate in the formation of pseudocleavage furrows, transient membranous invaginations between dividing nuclei. Embryos laid by homozygous sponge mothers lack these characteri...
Article
Full-text available
As part of a study of cytoskeletal proteins involved in Drosophila embryonic development, we have undertaken the molecular analysis of a 140-kD ATP-sensitive actin-binding protein (Miller, K. G., C. M. Field, and B. M. Alberts. 1989. J. Cell Biol. 109:2963-2975). Analysis of cDNA clones encoding this protein revealed that it represents a new class...
Article
The proteins that bind to actin filaments and microtubules play an important role in determining the structure and function of these filaments in eukaryotic cells. A number of approaches have been used to identify and characterize these proteins. Actin-binding proteins (ABPs) have been identified primarily by virtue of their ability to affect actin...
Article
Full-text available
By using F-actin affinity chromatography columns to select proteins solely by their ability to bind to actin filaments, we have identified and partially purified greater than 40 proteins from early Drosophila embryos. These proteins represent approximately 0.5% of the total protein present in soluble cell extracts, and 2 mg are obtained by chromato...
Article
By using F-actin affinity chromatography columns to select proteins solely by their ability to bind to actin filaments, we have identified and partially purified greater than 40 proteins from early Drosophila embryos. These proteins represent approximately 0.5% of the total protein present in soluble cell extracts, and 2 mg are obtained by chromato...
Article
We have developed stable and easy to use filamentous actin (F-actin) affinity-chromatography columns that selectively purify proteins that bind to actin filaments from cell extracts. Most traditional assays for actin-associated proteins screen for their effects on actin polymerization or actin filament crosslinking. Because our technique requires o...
Article
Full-text available
Three yeast actin-binding proteins were identified using yeast actin filaments as an affinity matrix. One protein appears to be a yeast myosin heavy chain; it is dissociated from actin filaments by ATP, it is similar in size (200 kD) to other myosins, and antibodies directed against Dictyostelium myosin heavy chain bind to it. Immunofluorescence ex...
Article
Full-text available
To determine the size and location of the mouse rDNA promoter, we constructed systematic series of deletion mutants approaching the initiation site from the 5' and 3' directions. These templates were transcribed in vitro under various conditions with S-100 and whole-cell extracts. Surprisingly, the size of the rDNA region that determines the level...
Article
To determine the size and location of the mouse rDNA promoter, we constructed systematic series of deletion mutants approaching the initiation site from the 5' and 3' directions. These templates were transcribed in vitro under various conditions with S-100 and whole-cell extracts. Surprisingly, the size of the rDNA region that determines the level...
Article
Excerpt We are attempting to use the early, single-cell Drosophila embryo as a model system to study two central biological problems. The first relates to the high degree of organization found in the intracellular compartment termed the cytosol (defined as the cytoplasm outside of membrane-bounded organelles). Cells apparently have the ability to p...
Article
Full-text available
The temperature-sensitive Drosophila mutation l(3)c21RRW630 disturbs oogenesis and imaginal disc development and has a maternal effect on embryogenesis. Two-dimensional gel electrophoretic analysis of protein synthesis in mutant tissue at a restrictive temperature shows that the synthesis of three proteins is elevated and the synthesis of three oth...
Chapter
From information presented in this volume and elsewhere, it is now clear that major control of eukaryotic gene expression is exerted at the transcriptional level, apparently largely at the initiation step. Consequently, much interest is being devoted to analyzing the transcriptional process, both in vivo and in vitro. Studies in manipulable systems...
Article
Full-text available
The in vitro initiation site for RNA polymerase I on the mouse rRNA gene was identified using a new method that is generally applicable to the study of other eukaryotic transcripts. First, the 5' end of mouse rRNA was located to an ApC . . . by high resolution S1 nuclease mapping. Dinucleotide primers were then used in transcription reactions to de...
Article
We have studied specific transcription by RNA polymerase I in vitro using a mouse tissue-culture cell extract and cloned mouse rDNA containing the transcription initiation region of a 45S rRNA gene. In vitro, transcription initiates at a unique site on the rDNA, which we have located using runoff transcription and S1 nuclease mapping. This site is...
Article
Full-text available
Using kinetoplast DNA networks as a substrate in a decatenation assay, we have purified to apparent homogeneity a type II DNA topoisomerase from HeLa cell nuclei. The most pure preparations contain a single polypeptide of 172,000 daltons as determined by sodium dodecyl sulfate-gel electrophoresis. The molecular weight of the native protein, based o...
Article
Two type I DNA topoisomerases have been purified to homogeneity from the nuclei of HeLa cells. One topoisomerase has a peptide molecular weight of 100,000 and the other, a molecular weight of 67,000. Several lines of evidence indicate that these two topoisomerases are closely related, (a) Both exhibit similar enzymatic activities on DNA. (b) The ch...
Article
Full-text available
Kinetoplast DNA is the mitochondrial DNA of trypanosomatids such as Crithidia fasciculata. This DNA is in the form of networks containing thousands of DNA circles which are apparently catenated (interlocked). Some topoisomerases, such as T4 phage topoisomerase and DNA gyrase, catalyze a decatenation of the networks to form individual covalently clo...

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