Functional analysis of the C-terminal domain in the Drosophila JIL-1 histone H3 kinase

Epigenetic regulation is an important process utilized by biological systems to control gene expression and organize development, with histone modification enzymes among the most important components in this regulatory network. The predominant interphase H3 Serine10 kinase in Drosophila is JIL-1, which plays an important role in regulating chromosome structure and modulating gene expression. The full-length JIL-1 protein is composed of four domains including a N-terminal domain (NTD), two kinase domains (KDI and KDII), and a unique C-terminal domain (CTD). In order to study the biological importance of these individual domains, a domain analysis research approach was taken and different functions of the JIL-1 kinase were dissected. This thesis will emphasize the significance of the CTD.;From a yeast-two-hybrid screening using the JIL-1 CTD as bait, lamin Dm0 was identified as a potential interaction partner. This interaction was further confirmed by molecular interaction analysis such as pull-down assays and co-immunoprecipitation experiments, and the interface for this interaction was mapped to the predicted globular region within JIL-1 CTD and the C-terminal tail in lamin Dm0. In the JIL-1z2/JIL-1h9 mutant devoid of the lamin interaction domain, we observed a distorted lamina structure in the nurse cells during oogenesis. In addition, genetic analysis in both viability assays and wm4 PEV (positional effect variegation) assays indicates that JIL-1 and lamin Dm0 counteract with each other.;Transgenic analysis using both the CFP-CTD and GFP-Delta-CTD transgenes indicates that the CTD is required for the proper chromosomal localization of JIL-1. Without the CTD, the truncated JIL-1 protein GFP-Delta-CTD loses its affinity for the chromosomes, and mainly localizes to the inter-chromosomal region. I identified a novel H3 binding motif within the CTD that may mediate the association between the CTD and chromatin. Rescue experiments using these transgenes also indicate that the CTD plays an important role in maintaining higher-order chromosome structure with the male X chromosome in particular.;In a PEV system where reporter genes are inserted in pericentric regions, loss of JIL-1 protein results in a dramatic reduction of reporter gene expression. This is consistent with the observation in JIL-1 loss-of-function mutants that the heterochromatin marker H3K9me2 mediated by the histone methyltransferase Su(var)3-9 spreads from the chromocenter to the chromosome arms; however, genetic analysis indicates that JIL-1 and Su(var)3-9 are likely to interact in a novel pathway that is largely independent of HP1. In contrast to the loss-of-function mutants the JIL-1Su(var)3-1 mutant, which carries an early stop codon that removes half of the CTD including both lamin and H3 interaction domains, dramatically increases the expression from the pericentric reporter genes and acts as a gain-of-function mutation in PEV.;In summary, the data above suggest a model in which the CTD of JIL-1 interacts with lamin Dm0 in Drosophila and this interaction is important for the integrity of nuclear lamina. In addition, the CTD can also interact with chromatin and may block the association between lamin and chromatin. The kinase activity of JIL-1 mediates the phosphorylation of H3S10, which can further open the chromatin, allowing the access of RNA Pol II and other transcription machineries and facilitating gene expression

Private Model Compression via Knowledge Distillation

The soaring demand for intelligent mobile applications calls for deploying powerful deep neural networks (DNNs) on mobile devices. However, the outstanding performance of DNNs notoriously relies on increasingly complex models, which in turn is associated with an increase in computational expense far surpassing mobile devices' capacity. What is worse, app service providers need to collect and utilize a large volume of users' data, which contain sensitive information, to build the sophisticated DNN models. Directly deploying these models on public mobile devices presents prohibitive privacy risk. To benefit from the on-device deep learning without the capacity and privacy concerns, we design a private model compression framework RONA. Following the knowledge distillation paradigm, we jointly use hint learning, distillation learning, and self learning to train a compact and fast neural network. The knowledge distilled from the cumbersome model is adaptively bounded and carefully perturbed to enforce differential privacy. We further propose an elegant query sample selection method to reduce the number of queries and control the privacy loss. A series of empirical evaluations as well as the implementation on an Android mobile device show that RONA can not only compress cumbersome models efficiently but also provide a strong privacy guarantee. For example, on SVHN, when a meaningful $(9.83,10^{-6})$-differential privacy is guaranteed, the compact model trained by RONA can obtain 20$\times$ compression ratio and 19$\times$ speed-up with merely 0.97% accuracy loss.Comment: Conference version accepted by AAAI'1

Pinning Cluster Synchronization in Linear Hybrid Coupled Delayed Dynamical Networks

The problem on cluster synchronization will be investigated for a class of delayed dynamical networks based on pinning control strategy. Through utilizing the combined convex technique and Kronecker product, two sufficient conditions can be derived to ensure the desired synchronization when the designed feedback controller is employed to each cluster. Moreover, the inner coupling matrices are unnecessarily restricted to be diagonal and the controller design can be converted into solving a series of linear matrix inequalities (LMIs), which greatly improve the present methods. Finally, two numerical examples are provided to demonstrate the effectiveness and reduced conservatism

Effects of Chilling on the Structure, Function and Development of Chloroplasts

Chloroplasts are the organelles that perform energy transformation in plants. The normal physiological functions of chloroplasts are essential for plant growth and development. Chilling is a common environmental stress in nature that can directly affect the physiological functions of chloroplasts. First, chilling can change the lipid membrane state and enzyme activities in chloroplasts. Then, the efficiency of photosynthesis declines, and excess reactive oxygen species (ROS) are produced. On one hand, excess ROS can damage the chloroplast lipid membrane; on the other hand, ROS also represent a stress signal that can alter gene expression in both the chloroplast and nucleus to help regenerate damaged proteins, regulate lipid homeostasis, and promote plant adaptation to low temperatures. Furthermore, plants assume abnormal morphology, including chlorosis and growth retardation, with some even exhibiting severe necrosis under chilling stress. Here, we review the response of chloroplasts to low temperatures and focus on photosynthesis, redox regulation, lipid homeostasis, and chloroplast development to elucidate the processes involved in plant responses and adaptation to chilling stress

Histone H3S10 phosphorylation by the JIL-1 kinase in pericentric heterochromatin and on the fourth chromosome creates a composite H3S10phK9me2 epigenetic mark

The JIL-1 kinase mainly localizes to euchromatic interband regions of polytene chromosomes and is the kinase responsible for histone H3S10 phosphorylation at interphase in Drosophila. However, recent findings raised the possibility that the binding of some H3S10ph antibodies may be occluded by the H3K9me2 mark obscuring some H3S10 phosphorylation sites. Therefore, we have characterized an antibody to the epigenetic H3S10phK9me2 double mark as well as three commercially available H3S10ph antibodies. The results showed that for some H3S10ph antibodies their labeling indeed can be occluded by the concomitant presence of the H3K9me2 mark. Furthermore, we demonstrate that the double H3S10phK9me2 mark is present in pericentric heterochromatin as well as on the fourth chromosome of wild-type polytene chromosomes but not in preparations from JIL-1 or Su(var)3-9 null larvae. Su(var)3-9 is a methyltransferase mediating H3K9 dimethylation. Furthermore, the H3S10phK9me2 labeling overlapped with that of the non-occluded H3S10ph antibodies as well as with H3K9me2 antibody labeling. Interestingly, when a Lac-I-Su(var)3-9 transgene is overexpressed, it upregulates H3K9me2 dimethylation on the chromosome arms creating extensive ectopic H3S10phK9me2 marks suggesting that the H3K9 dimethylation occurred at euchromatic H3S10ph sites. This is further supported by the finding that under these conditions euchromatic H3S10ph labeling by the occluded antibodies was abolished. Thus, our findings indicate a novel role for the JIL-1 kinase in epigenetic regulation of heterochromatin in the context of the chromocenter and fourth chromosome by creating a composite H3S10phK9me2 mark together with the Su(var)3-9 methyltransferase

RNA polymerase II-mediated transcription at active loci does not require histone H3S10 phosphorylation in Drosophila

JIL-1 is the major kinase controlling the phosphorylation state of histone H3S10 at interphase in Drosophila. In this study, we used three different commercially available histone H3S10 phosphorylation antibodies, as well as an acid-free polytene chromosome squash protocol that preserves the antigenicity of the histone H3S10 phospho-epitope, to examine the role of histone H3S10 phosphorylation in transcription under both heat shock and non-heat shock conditions. We show that there is no redistribution or upregulation of JIL-1 or histone H3S10 phosphorylation at transcriptionally active puffs in such polytene squash preparations after heat shock treatment. Furthermore, we provide evidence that heat shock-induced puffs in JIL-1 null mutant backgrounds are strongly labeled by antibody to the elongating form of RNA polymerase II (Pol IIoser2), indicating that Pol IIoser2 is actively involved in heat shock-induced transcription in the absence of histone H3S10 phosphorylation. This is supported by the finding that there is no change in the levels of Pol IIoser2 in JIL-1 null mutant backgrounds compared with wild type. mRNA from the six genes that encode the major heat shock protein in Drosophila, Hsp70, is transcribed at robust levels in JIL-1 null mutants, as directly demonstrated by qRT-PCR. Taken together, these data are inconsistent with the model that Pol II-dependent transcription at active loci requires JIL-1-mediated histone H3S10 phosphorylation, and instead support a model in which transcriptional defects in the absence of histone H3S10 phosphorylation are a result of structural alterations of chromatin