Craig D. Kaplan
Texas A&M University
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Featured researches published by Craig D. Kaplan.
Chemical Reviews | 2009
Craig D. Kaplan; Jerry Kaplan
Iron is an essential element for all eukaryotes. The facile ability of iron to gain and lose electrons permits its participation in a wide variety of oxidation-reduction reactions. Further, the ability of iron as a component of heme to bind oxygen makes it indispensable as an oxygen carrier and sensor, particularly in vertebrates where the size of the organism provides a barrier to simple oxygen diffusion requiring specialized oxygen transport. Iron is found as a prosthetic group on proteins in various forms: elemental iron, oxoiron or oxoiron-zinc, heme, and iron-sulfur clusters. While the diversity of iron-containing proteins and the reactions they participate in highlights the usefulness of iron in biochemical reactions, iron is problematic for two reasons. First, the same facile property of electron gain and loss that permits iron to participate in oxidation-reduction reactions permits iron to donate electrons to oxygen or hydrogen peroxide, generating toxic oxygen radicals, superoxide anion and hydroxyl radical. The existence of these oxygen radicals has led to the evolution of enzyme systems capable of either preventing or removing oxygen radicals once formed. Loss of these enzymes, such as superoxide dismutase, leads to increased cellular damage and compromised growth. Because iron can be toxic, all iron acquisition systems are highly regulated in order to restrict the concentration of iron within biological fluids, extending from the cytosol of yeast to the plasma of vertebrates. Furthermore, regulation of iron acquisition is one of the most critical steps in maintaining iron homeostasis because eukaryotes do not have regulated mechanisms of iron egress. The second reason that iron is problematic is that bioavailable iron is scarce. Iron is the fourth most abundant element on earth but is not uniformly distributed. Iron is scarce both because there are vast geographical areas, such as the oceans, which are iron-poor and because iron, when abundant, is found in a biologically inaccessible form. Iron’s proclivity to react with oxygen has resulted in the predominant form of iron being the insoluble ferric hydroxide. Great investment of cellular resources is required to make iron bioavailable. Even with such effort, the external concentration of iron is often limiting, leading to cellular responses that “triage” iron to biochemical pathways that most require it. We have come to understand that not only is the acquisition and storage of iron highly regulated, but the processes that divert iron among intracellular biochemical pathways are also * Corresponding author. † Texas A&M University. ‡ University of Utah. Craig D. Kaplan obtained a B.S. in Biology and Latin Language and Literature from the University of Michigan, Ann Arbor, Michigan, in 1995, and a Ph.D. in Genetics from Harvard University in 2003 under the supervision of Dr. Fred Winston at Harvard Medical School. His Ph.D. dissertation focused on the regulation of chromatin structure and transcription by the RNA Polymerase II elongation factor Spt6. Much of this manuscript was written while he was a postdoctoral fellow with Dr. Roger Kornberg at the Stanford University School of Medicine, investigating the mechanisms of RNA Polymerase II elongation control, both innate and factor-mediated. He has recently joined the faculty in the Department of Biochemistry and Biophysics, at Texas A&M University, as an assistant professor. Chem. Rev. 2009, 109, 4536–4552 4536
Proceedings of the National Academy of Sciences of the United States of America | 2012
Matthew H. Larson; Jing Zhou; Craig D. Kaplan; Murali Palangat; Roger D. Kornberg; Robert Landick; Steven M. Block
During transcription, RNA polymerase II (RNAPII) must select the correct nucleotide, catalyze its addition to the growing RNA transcript, and move stepwise along the DNA until a gene is fully transcribed. In all kingdoms of life, transcription must be finely tuned to ensure an appropriate balance between fidelity and speed. Here, we used an optical-trapping assay with high spatiotemporal resolution to probe directly the motion of individual RNAPII molecules as they pass through each of the enzymatic steps of transcript elongation. We report direct evidence that the RNAPII trigger loop, an evolutionarily conserved protein subdomain, serves as a master regulator of transcription, affecting each of the three main phases of elongation, namely: substrate selection, translocation, and catalysis. Global fits to the force-velocity relationships of RNAPII and its trigger loop mutants support a Brownian ratchet model for elongation, where the incoming NTP is able to bind in either the pre- or posttranslocated state, and movement between these two states is governed by the trigger loop. Comparison of the kinetics of pausing by WT and mutant RNAPII under conditions that promote base misincorporation indicate that the trigger loop governs fidelity in substrate selection and mismatch recognition, and thereby controls aspects of both transcriptional accuracy and rate.
Cell | 2013
Hannes Braberg; Huiyan Jin; Erica A. Moehle; Yujia A. Chan; Shuyi Wang; Michael Shales; Joris J. Benschop; John H. Morris; Chenxi Qiu; Fuqu Hu; Leung K. Tang; J.S. Fraser; Frank C. P. Holstege; Philip Hieter; Christine Guthrie; Craig D. Kaplan; Nevan J. Krogan
RNA polymerase II (RNAPII) lies at the core of dynamic control of gene expression. Using 53 RNAPII point mutants, we generated a point mutant epistatic miniarray profile (pE-MAP) comprising ∼60,000 quantitative genetic interactions in Saccharomyces cerevisiae. This analysis enabled functional assignment of RNAPII subdomains and uncovered connections between individual regions and other protein complexes. Using splicing microarrays and mutants that alter elongation rates in vitro, we found an inverse relationship between RNAPII speed and in vivo splicing efficiency. Furthermore, the pE-MAP classified fast and slow mutants that favor upstream and downstream start site selection, respectively. The striking coordination of polymerization rate with transcription initiation and splicing suggests that transcription rate is tuned to regulate multiple gene expression steps. The pE-MAP approach provides a powerful strategy to understand other multifunctional machines at amino acid resolution.
PLOS Genetics | 2012
Craig D. Kaplan; Huiyan Jin; Ivan Liang Zhang; Andrey Belyanin
Structural and biochemical studies have revealed the importance of a conserved, mobile domain of RNA Polymerase II (Pol II), the Trigger Loop (TL), in substrate selection and catalysis. The relative contributions of different residues within the TL to Pol II function and how Pol II activity defects correlate with gene expression alteration in vivo are unknown. Using Saccharomyces cerevisiae Pol II as a model, we uncover complex genetic relationships between mutated TL residues by combinatorial analysis of multiply substituted TL variants. We show that in vitro biochemical activity is highly predictive of in vivo transcription phenotypes, suggesting direct relationships between phenotypes and Pol II activity. Interestingly, while multiple TL residues function together to promote proper transcription, individual residues can be separated into distinct functional classes likely relevant to the TL mechanism. In vivo, Pol II activity defects disrupt regulation of the GTP-sensitive IMD2 gene, explaining sensitivities to GTP-production inhibitors, but contrasting with commonly cited models for this sensitivity in the literature. Our data provide support for an existing model whereby Pol II transcriptional activity provides a proxy for direct sensing of NTP levels in vivo leading to IMD2 activation. Finally, we connect Pol II activity to transcription start site selection in vivo, implicating the Pol II active site and transcription itself as a driver for start site scanning, contravening current models for this process.
Molecular Cell | 2015
Célia Jeronimo; Shinya Watanabe; Craig D. Kaplan; Craig L. Peterson; François Robert
H2A.Z is a highly conserved histone variant involved in several key nuclear processes. It is incorporated into promoters by SWR-C-related chromatin remodeling complexes, but whether it is also actively excluded from non-promoter regions is not clear. Here we provide genomic and biochemical evidence that the RNA polymerase II (RNA Pol II) elongation-associated histone chaperones FACT and Spt6 both contribute to restricting H2A.Z from intragenic regions. In the absence of FACT or Spt6, the lack of efficient nucleosome reassembly coupled to pervasive incorporation of H2A.Z by mislocalized SWR-C alters chromatin composition and contributes to cryptic initiation. Therefore, chaperone-mediated H2A.Z confinement is crucial for restricting the chromatin signature of gene promoters that otherwise may license or promote cryptic transcription.
Nature | 2016
Jeremy G. Bird; Yu Zhang; Yuan Tian; Natalya Panova; Ivan Barvík; Landon Greene; Min Liu; Brian Buckley; Libor Krásný; Jeehiun K. Lee; Craig D. Kaplan; Richard H. Ebright; Bryce E. Nickels
The chemical nature of the 5′ end of RNA is a key determinant of RNA stability, processing, localization and translation efficiency, and has been proposed to provide a layer of ‘epitranscriptomic’ gene regulation. Recently it has been shown that some bacterial RNA species carry a 5′-end structure reminiscent of the 5′ 7-methylguanylate ‘cap’ in eukaryotic RNA. In particular, RNA species containing a 5′-end nicotinamide adenine dinucleotide (NAD+) or 3′-desphospho-coenzyme A (dpCoA) have been identified in both Gram-negative and Gram-positive bacteria. It has been proposed that NAD+, reduced NAD+ (NADH) and dpCoA caps are added to RNA after transcription initiation, in a manner analogous to the addition of 7-methylguanylate caps. Here we show instead that NAD+, NADH and dpCoA are incorporated into RNA during transcription initiation, by serving as non-canonical initiating nucleotides (NCINs) for de novo transcription initiation by cellular RNA polymerase (RNAP). We further show that both bacterial RNAP and eukaryotic RNAP II incorporate NCIN caps, that promoter DNA sequences at and upstream of the transcription start site determine the efficiency of NCIN capping, that NCIN capping occurs in vivo, and that NCIN capping has functional consequences. We report crystal structures of transcription initiation complexes containing NCIN-capped RNA products. Our results define the mechanism and structural basis of NCIN capping, and suggest that NCIN-mediated ‘ab initio capping’ may occur in all organisms.
Journal of Biological Chemistry | 2010
Man-Hee Suh; Peter Meyer; Meigang Gu; Ping Ye; Mincheng Zhang; Craig D. Kaplan; Christopher D. Lima; Jianhua Fu
RNA capping enzyme (CE) is recruited specifically to RNA polymerase II (Pol II) transcription sites to facilitate cotranscriptional 5′-capping of pre-mRNA and other Pol II transcripts. The current model to explain this specific recruitment of CE to Pol II as opposed to Pol I and Pol III rests on the interaction between CE and the phosphorylated C-terminal domain (CTD) of Pol II largest subunit Rpb1 and more specifically between the CE nucleotidyltransferase domain and the phosphorylated CTD. Through biochemical and diffraction analyses, we demonstrate the existence of a distinctive stoichiometric complex between CE and the phosphorylated Pol II (Pol IIO). Analysis of the complex revealed an additional and unexpected polymerase-CE interface (PCI) located on the multihelical Foot domain of Rpb1. We name this interface PCI1 and the previously known nucleotidyltransferase/phosphorylated CTD interface PCI2. Although PCI1 and PCI2 individually contribute to only weak interactions with CE, a dramatically stabilized and stoichiometric complex is formed when PCI1 and PCI2 are combined in cis as they occur in an intact phosphorylated Pol II molecule. Disrupting either PCI1 or PCI2 by alanine substitution or deletion diminishes CE association with Pol II and causes severe growth defects in vivo. Evidence from manipulating PCI1 indicates that the Foot domain contributes to the specificity in CE interaction with Pol II as opposed to Pol I and Pol III. Our results indicate that the dual interface based on combining PCI1 and PCI2 is required for directing CE to Pol II elongation complexes.
Proceedings of the National Academy of Sciences of the United States of America | 2014
Volker Schweikhard; Cong Meng; Kenji Murakami; Craig D. Kaplan; Roger D. Kornberg; Steven M. Block
Significance In higher organisms, DNA is bound to proteins and tightly packed within the nucleus, leaving only certain regions accessible for gene expression. As the enzyme RNA polymerase II (RNAPII) travels along the DNA template synthesizing RNA, it must contend with forces generated by various obstacles, and some of these forces are known to produce transient pausing and even transcriptional arrest. How do organisms deal with such problems? Using optical-trapping technology, we performed a single-molecule study of RNAPII interactions with transcription factors TFIIS and TFIIF, which are involved in modulating and regulating transcriptional elongation. By applying controlled loads to RNAPII and combinations of these factors, we learned about the mechanisms by which pausing and arrest are overcome. Recent evidence suggests that transcript elongation by RNA polymerase II (RNAPII) is regulated by mechanical cues affecting the entry into, and exit from, transcriptionally inactive states, including pausing and arrest. We present a single-molecule optical-trapping study of the interactions of RNAPII with transcription elongation factors TFIIS and TFIIF, which affect these processes. By monitoring the response of elongation complexes containing RNAPII and combinations of TFIIF and TFIIS to controlled mechanical loads, we find that both transcription factors are independently capable of restoring arrested RNAPII to productive elongation. TFIIS, in addition to its established role in promoting transcript cleavage, is found to relieve arrest by a second, cleavage-independent mechanism. TFIIF synergistically enhances some, but not all, of the activities of TFIIS. These studies also uncovered unexpected insights into the mechanisms underlying transient pauses. The direct visualization of pauses at near-base-pair resolution, together with the load dependence of the pause-entry phase, suggests that two distinct mechanisms may be at play: backtracking under forces that hinder transcription and a backtrack-independent activity under assisting loads. The measured pause lifetime distributions are inconsistent with prevailing views of backtracking as a purely diffusive process, suggesting instead that the extent of backtracking may be modulated by mechanisms intrinsic to RNAPII. Pauses triggered by inosine triphosphate misincorporation led to backtracking, even under assisting loads, and their lifetimes were reduced by TFIIS, particularly when aided by TFIIF. Overall, these experiments provide additional insights into how obstacles to transcription may be overcome by the concerted actions of multiple accessory factors.
Biochimica et Biophysica Acta | 2013
Craig D. Kaplan
Transcription by RNA polymerase II (Pol II), and all RNA polymerases for that matter, may be understood as comprising two cycles. The first cycle relates to the basic mechanism of the transcription process wherein Pol II must select the appropriate nucleoside triphosphate (NTP) substrate complementary to the DNA template, catalyze phosphodiester bond formation, and translocate to the next position on the DNA template. Performing this cycle in an iterative fashion allows the synthesis of RNA chains that can be over one million nucleotides in length in some larger eukaryotes. Overlaid upon this enzymatic cycle, transcription may be divided into another cycle of three phases: initiation, elongation, and termination. Each of these phases has a large number of associated transcription factors that function to promote or regulate the gene expression process. Complicating matters, each phase of the latter transcription cycle are coincident with cotranscriptional RNA processing events. Additionally, transcription takes place within a highly dynamic and regulated chromatin environment. This chromatin environment is radically impacted by active transcription and associated chromatin modifications and remodeling, while also functioning as a major platform for Pol II regulation. This review will focus on our basic knowledge of the Pol II transcription mechanism, and how altered Pol II activity impacts gene expression in vivo in the model eukaryote Saccharomyces cerevisiae. This article is part of a Special Issue entitled: RNA Polymerase II Transcript Elongation.
Proceedings of the National Academy of Sciences of the United States of America | 2012
Kenji Murakami; Brian J. Gibbons; Ralph E. Davis; Shigeki Nagai; Xin Liu; Philip J. J. Robinson; Tinghe Wu; Craig D. Kaplan; Roger D. Kornberg
General transcription factor TFIIH, previously described as a 10-subunit complex, is essential for transcription and DNA repair. An eleventh subunit now identified, termed Tfb6, exhibits 45% sequence similarity to human nuclear mRNA export factor 5. Tfb6 dissociates from TFIIH as a heterodimer with the Ssl2 subunit, a DNA helicase that drives promoter melting for the initiation of transcription. Tfb6 does not, however, dissociate Ssl2 from TFIIH in the context of a fully assembled transcription preinitiation complex. Our findings suggest a dynamic state of Ssl2, allowing its engagement in multiple cellular processes.