Novel Mekk1-p53 mediated transcriptional regulation: mechanism and genome wide st

新型 Mekk1-p53 介导的转录调控：机制和全基因组范围

• 批准号: 7883065
• 负责人: M. Rafiq Islam
• 金额: $ 20.88万
• 依托单位: NORTHWEST MISSOURI STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-15 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7883065
• 关键词: Academy; Affect; Apoptosis; Binding; Binding Sites; Biochemical; Biochemistry; Biological; Biomedical Research; Cell Nucleus; Cell physiology; Cells; Cellular biology; Collaborations; Cytoplasm; Degradation Pathway; Deletion Mutation; Educational Curriculum; Ensure; Environment; Exposure to; Funding; Future; Genes; Genetic Transcription; Goals; Housing; Immunoprecipitation; Infusion procedures; Institution; Kansas; Learning; MAP Kinase Kinase Kinase; MAPK8 gene; Mediating; Medical center; Membrane Proteins; Methodology; Missouri; Mitogen-Activated Protein Kinase Kinases; Mitogen-Activated Protein Kinases; Molecular; Nuclear; Nuclear Translocation; Pathway interactions; Phosphorylation; Phosphotransferases; Polycystic Kidney Diseases; Protein Sequence Analysis; Protein p53; Proteins; Regulation; Reporting; Research; Research Personnel; Research Training; SAPK; Science; Signal Transduction; Site; Students; TP53 gene; Techniques; Therapeutic; Training; Transcriptional Regulation; Universities; career; experience; gene repression; genome-wide; high school; improved; novel; overexpression; programs; promoter; public health relevance; transcription factor

DESCRIPTION (provided by applicant): MAPK (mitogen-activated protein kinase) cascades are involved in a variety of cellular functions including proliferation, differentiation, and apoptosis. Upon activation by any of a number of mechanisms, Mekk (a MAPK kinase kinase) activates Mkk (a MAPKK or Sek), which in turn activates SAPK (a MAPK). The MAPK is then translocated from the cytoplasm to the nucleus, where it directly regulates the activity of transcription factors controlling the expression of a number of genes. Recently, we observed Mekk1 and its constitutively active fragment CA-Mekk1 (1174-1493 aa) were translocated to the nucleus when overexpressed in HEK293T, COS, and HCT cells and affected transcription of the Pkd1 (Polycystic kidney disease) gene and possibly other genes via the anti-tumor protein, p53. As such, we have begun to unravel a novel mechanism of transcriptional regulation. Thus, with a broad goal to understand the novel mechanism of Mekk1-p53 mediated transcriptional regulation of the Pkd1 and other genes, we propose to accomplish the following four Specific Aims: Aim 1: Identify fragment(s) of Mekk1 translocating to the nucleus and determine their mechanism of nuclear translocation. Aim 2: Characterize the p53-Mekk1 binding mechanism by: (a) identifying the binding site if it is direct, or (b) identifying the intermediate partners, if it is indirect. Aim 3: Investigate the mechanism of p53 elevation. Aim 4: Perform ChIP-Seq analysis to identify genome-wide binding sites for Mekk1, p53 and both. We will use a variety of biochemical and cellular/molecular biological approaches as well as the latest cutting edge technique, ChIP-Seq. These studies will not only improve our understanding of transcriptional regulation by both p53 and Mekk1, but may also uncover the mechanism of the Mekk1-p53 pathway. Thus, the answers sought should have both fundamental significances and should provide information leading to future therapeutic approaches. The research will engage undergraduates at Northwest Missouri State University (NWMSU), as well as talented high school students in the Missouri Academy of Science, Math, and Computing (MASMC) housed on the Northwest campus. Training received in this project will prepare students for entry into science programs and/or scientific professional careers. Thus, the project will (i) provide undergraduate and talented high school students benefits of exposure to and participation in biomedical research, (ii) strengthen the research environment at NWMSU, and (iii) give the PI an opportunity to continue meritorious research. Furthermore, collaborations with investigators at a research-intensive institution (University of Kansas Medical Center) will ensure the infusion of the latest methodology and information into the biochemistry/cell biology curricula. Thus, the project will advance discovery and understanding while also promoting research, training and learning. PUBLIC HEALTH RELEVANCE: Recently, we unraveled a novel mechanism in which nuclear translocation of Mekk1 and its interaction with p53 leads to transcriptional repression of PKD1, independent of the MAPK phosphorylation cascade. The mechanism was previously unknown. This proposal will study the mechanism in detail and will seek to determine how Mekk1, a membrane-associated protein, translocates to the nucleus and interacts with p53; and will utilize cutting-edge genome-wide approaches to identify other genes that might be under regulation of this Mekk1-p53 pathway.

描述（由申请人提供）：MAPK（促分裂原活化蛋白激酶）级联参与多种细胞功能，包括增殖、分化和凋亡。在通过多种机制中的任一种激活后，Mekk（MAPK激酶激酶）激活Mkk（MAPKK或Sek），Mkk又激活SAPK（MAPK）。然后MAPK从细胞质转移到细胞核，在那里它直接调节控制许多基因表达的转录因子的活性。最近，我们观察到Mekk 1及其组成型活性片段CA-Mekk 1（1174-1493 aa）在HEK 293 T，COS和HCT细胞中过表达时易位到细胞核，并通过抗肿瘤蛋白p53影响Pkd 1（多囊肾病）基因和可能的其他基因的转录。因此，我们已经开始解开一个新的转录调控机制。因此，为了了解Mekk 1-p53介导的Pkd 1和其他基因转录调控的新机制，我们提出了以下四个具体目标：目标1：鉴定Mekk 1易位到细胞核的片段并确定其核易位机制。目标二：通过以下方式表征p53-Mekk 1结合机制：（a）如果是直接结合，则鉴定结合位点;或（B）如果是间接结合，则鉴定中间伴侣。目的3：探讨p53升高的机制。目的4：进行ChIP-Seq分析，以确定Mekk 1，p53和两者的全基因组结合位点。我们将使用各种生物化学和细胞/分子生物学方法以及最新的尖端技术ChIP-Seq。这些研究不仅将提高我们对p53和Mekk 1的转录调控的理解，而且还可能揭示Mekk 1-p53通路的机制。因此，寻求的答案应该具有根本意义，并应该提供导致未来治疗方法的信息。这项研究将吸引西北密苏里州州立大学（NWMSU）的本科生，以及西北校区内密苏里州科学、数学和计算学院（MASMC）的优秀高中生。在本项目中接受的培训将为学生进入科学课程和/或科学专业生涯做好准备。因此，该项目将（一）提供本科生和有才华的高中学生接触和参与生物医学研究的好处，（二）加强在NWMSU的研究环境，以及（三）给PI一个机会，继续有价值的研究。此外，与研究密集型机构（堪萨斯大学医学中心）的研究人员合作，将确保将最新的方法和信息注入生物化学/细胞生物学课程。因此，该项目将促进发现和理解，同时也促进研究、培训和学习。 公共卫生关系：最近，我们揭示了一种新的机制，其中Mekk 1的核转位及其与p53的相互作用导致PKD 1的转录抑制，独立于MAPK磷酸化级联反应。其机制以前是未知的。该提案将详细研究该机制，并将寻求确定Mekk 1（一种膜相关蛋白）如何易位到细胞核并与p53相互作用;并将利用尖端的全基因组方法来识别可能受Mekk 1-p53通路调控的其他基因。

项目成果

Nuclear Localization Signal and p53 Binding Site in MAP/ERK Kinase Kinase 1 (MEKK1).

• DOI: 10.1002/jcb.25238
• 发表时间: 2015-12
• 期刊: Journal of cellular biochemistry
• 影响因子: 4
• 作者: Chipps E;Protzman A;Muhi MZ;Ando S;Calvet JP;Islam MR
• 通讯作者: Islam MR

Regulation of HFE expression by poly(ADP-ribose) polymerase-1 (PARP1) through an inverted repeat DNA sequence in the distal promoter.

• DOI: 10.1016/j.bbagrm.2013.10.002
• 发表时间: 2013-12
• 期刊: BIOCHIMICA ET BIOPHYSICA ACTA-GENE REGULATORY MECHANISMS
• 影响因子: 4.7
• 作者: Pelham, Christopher;Jimenez, Tamara;Rodova, Marianna;Rudolph, Angela;Chipps, Elizabeth;Islam, M. Rafiq
• 通讯作者: Islam, M. Rafiq

Measuring h-index and scholarly productivity in academic dermatology in Canada.

• DOI: 10.1007/s11192-022-04589-y
• 发表时间: 2023-02
• 期刊: SCIENTOMETRICS
• 影响因子: 3.9
• 作者: Azar, Marleine;Lagace, Francois;Muntyanu, Anastasiya;Netchiporouk, Elena;Zhou, Youwen;Lynde, Charles;Moreau, Linda;Mathieu, Steve;Sasseville, Denis;Asiniwasis, Rachel;Shear, Neil H.;Gniadecki, Robert;Rahme, Elham;Litvinov, Ivan V.
• 通讯作者: Litvinov, Ivan V.