scholarly journals Protein Deacetylase CobB Interplays with c-di-GMP

2018 ◽  
Author(s):  
Zhaowei Xu ◽  
Hainan Zhang ◽  
Xingrun Zhang ◽  
Chengxi Liu ◽  
Hewei Jiang ◽  
...  
Keyword(s):  
Feedback Loop ◽  
Dna Supercoiling ◽  
Bacterial Motility ◽  
Mass Spectrometry Analysis ◽  
Diguanylate Cyclase ◽  
Major Site ◽  
Spectrometry Analysis ◽  
Secondary Messenger ◽  
Family Protein ◽  
Protein Deacetylase

AbstractAs a ubiquitous bacterial secondary messenger, c-di-GMP plays key regulatory roles in processes such as bacterial motility and transcription regulation. CobB is the Sir2 family protein deacetylase that controls energy metabolism, chemotaxis and DNA supercoiling in many bacteria. Using anE.coliproteome microarray, we found that c-di-GMP strongly binds to CobB. Protein deacetylation assays showed that c-di-GMP inhibits CobB activity and thereby modulates the biogenesis of acetyl-CoA. Through mutagenesis studies, residues R8, R17 and E21 of CobB were shown to be required for c-di-GMP binding. Next, we found that CobB is an effective deacetylase of YdeH, a major diguanylate cyclase (DGC) ofE.colithat is endogenously acetylated. Mass spectrometry analysis identified YdeH K4 as the major site of acetylation, and it could be deacetylated by CobB. Interestingly, deacetylation of YdeH enhances its stability and cyclase activity in c-di-GMP production. Thus, our work establishes a novel negative feedback loop linking c-di-GMP biogenesis and CobB-mediated protein deacetylation.

scholarly journals CEP131 Abrogates CHK1 Inhibitor-Induced Replication Defects and Is Associated with Unfavorable Outcome in Neuroblastoma

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Kiyohiro Ando ◽  
Verna Cázares-Ordoñez ◽  
Makoto Makishima ◽  
Atsushi Yokoyama ◽  
Yusuke Suenaga ◽  
...  
Keyword(s):  
Dna Damage ◽  
Therapeutic Strategy ◽  
Neuroblastoma Cell ◽  
Mass Spectrometry Analysis ◽  
Spectrometry Analysis ◽  
Checkpoint Kinase 1 ◽  
Family Protein ◽  
Damage Response ◽  
Chk1 Inhibitor ◽  
Associated Family

Checkpoint kinase 1 (CHK1) plays a key role in genome surveillance and integrity throughout the cell cycle. Selective inhibitors of CHK1 (CHK1i) are undergoing clinical evaluation for various human malignancies, including neuroblastoma. Recently, we reported that CHK1i, PF-477736, induced a p53-mediated DNA damage response. As a result, the cancer cells were able to repair DNA damage and became less sensitive to CHK1i. In this study, we discovered that PF-477736 increased expression of MDM2 oncogene along with CHK1i-induced replication defects in neuroblastoma NB-39-nu cells. A mass spectrometry analysis of protein binding to MDM2 in the presence of CHK1i identified the centrosome-associated family protein 131 (CEP131), which was correlated with unfavorable prognosis of neuroblastoma patients. We revealed that MDM2 was associated with CEP131 protein degradation, whereas overexpression of CEP131 accelerated neuroblastoma cell growth and exhibited resistance to CHK1i-induced replication defects. Thus, these findings may provide a future therapeutic strategy against centrosome-associated oncogenes involving CEP131 as a target in neuroblastoma.

scholarly journals Roles of paxillin family members in adhesion and ECM degradation coupling at invadosomes

2016 ◽  
Vol 213 (5) ◽  
pp. 585-599 ◽  
Author(s):  
Christos Petropoulos ◽  
Christiane Oddou ◽  
Anouk Emadali ◽  
Edwige Hiriart-Bryant ◽  
Cyril Boyault ◽  
...  
Keyword(s):  
Family Members ◽  
Janus Kinase ◽  
Mass Spectrometry Analysis ◽  
Spectrometry Analysis ◽  
Adhesion Sites ◽  
Lim Domains ◽  
Janus Kinase 1 ◽  
Family Protein ◽  
Functional Complex ◽  
Ecm Degradation

Invadosomes are acto-adhesive structures able to both bind the extracellular matrix (ECM) and digest it. Paxillin family members—paxillin, Hic-5, and leupaxin—are implicated in mechanosensing and turnover of adhesion sites, but the contribution of each paxillin family protein to invadosome activities is unclear. We use genetic approaches to show that paxillin and Hic-5 have both redundant and distinctive functions in invadosome formation. The essential function of paxillin-like activity is based on the coordinated activity of LD motifs and LIM domains, which support invadosome assembly and morphology, respectively. However, paxillin preferentially regulates invadosome assembly, whereas Hic-5 regulates the coupling between ECM degradation and acto-adhesive functions. Mass spectrometry analysis revealed new partners that are important for paxillin and Hic-5 specificities: paxillin regulates the acto-adhesive machinery through janus kinase 1 (JAK1), whereas Hic-5 controls ECM degradation via IQGAP1. Integrating the redundancy and specificities of paxillin and Hic-5 in a functional complex provides insights into the coupling between the acto-adhesive and ECM-degradative machineries in invadosomes.

scholarly journals Structure of a Novel Bacterial c-GMP Binding Protein

2014 ◽  
Vol 70 (a1) ◽  
pp. C1637-C1637
Author(s):  
Shan-Ho Chou
Keyword(s):  
Xanthomonas Campestris ◽  
Binding Protein ◽  
Isothermal Calorimetry ◽  
Receptor Protein ◽  
Diguanylate Cyclase ◽  
Amino Acid Residues ◽  
Rhodospirillum Centenum ◽  
Secondary Messenger ◽  
Family Protein ◽  
Cgmp Signaling

cAMP is an important secondary messenger molecule widely distributed across all living kingdoms, whereas cGMP is generally considered to be restricted to eukaryotes. Recently, solid evidences for cGMP signaling in Rhodospirillum centenum have been provided, and it is proposed that cGMP could also be adapted to deliver messages to diverse outputs via unknown mechanisms. While the structures and functions of binding between cAMP and its receptor protein CRP have been well studied in the past, currently no structure of prokaryotic cGMP-binding protein complex is known. Here we report the first determination of a cGMP-receptor crystal structure from the plant pathogen Xanthomonas campestris (Xcc) to a resolution of 2.2 Å. The new cGMP receptor Xcc0249 is found to belong to the CRP/FNR family protein containing both a cyclic-Nucleotide Binding Domain (cNBD) and a GGDEF domains, and exhibits strong cGMP binding and diguanylate cyclase activities. Mutations of crucial amino acid residues responsible for cGMP binding to Xcc0249 are found to significantly reduce the biofilm formation and virulence in Xcc. Isothermal calorimetry (ITC) measurements demonstrate that Xcc0249 can bind preferentially to cGMP with a much stronger affinity (KD: 2.93E-7) than cAMP (KD: 1.79E-5). cGMP binding to Xcc0249 is also found to enhance the GGDEF diguanylate cyclase activity, implying a broader functional role of cGMP and a possible linkage between the cGMP and c-di-GMP interaction networks in bacteria. References

scholarly journals Control of Acetyl-Coenzyme A Synthetase (AcsA) Activity by Acetylation/Deacetylation without NAD+ Involvement in Bacillus subtilis

2006 ◽  
Vol 188 (15) ◽  
pp. 5460-5468 ◽  
Author(s):  
Jeffrey G. Gardner ◽  
Frank J. Grundy ◽  
Tina M. Henkin ◽  
Jorge C. Escalante-Semerena
Keyword(s):  
Bacillus Subtilis ◽  
Coenzyme A ◽  
Mass Spectrometry Analysis ◽  
Acetyl Coenzyme A ◽  
Spectrometry Analysis ◽  
Acetyl Coenzyme ◽  
Protein Acetyltransferase ◽  
Protein Deacetylase

ABSTRACT Posttranslational modification is an efficient mechanism for controlling the activity of structural proteins, gene expression regulators, and enzymes in response to rapidly changing physiological conditions. Here we report in vitro and in vivo evidence that the acuABC operon of the gram-positive soil bacterium Bacillus subtilis encodes a protein acetyltransferase (AcuA) and a protein deacetylase (AcuC), which may control the activity of acetyl-coenzyme A (CoA) synthetase (AMP-forming, AcsA) in this bacterium. Results from in vitro experiments using purified proteins show that AcsA is a substrate for the acetyl-CoA-dependent AcuA acetyltransferase. Mass spectrometry analysis of a tryptic digest of acetylated AcsA (AcsAAc) identified residue Lys549 as the sole modification site in the protein. Unlike sirtuins, the AcuC protein did not require NAD+ as cosubstrate to deacetylate AcsAAc. The function of the putative AcuB protein remains unknown.

scholarly journals Identification of Phosphorylated Amino Acids in Human TNRC6A C-Terminal Region and Their Effects on the Interaction with the CCR4-NOT Complex

Genes ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 271
Author(s):  
Fusako Munakata ◽  
Masataka Suzawa ◽  
Kumiko Ui-Tei
Keyword(s):  
Rna Silencing ◽  
Terminal Region ◽  
Scaffold Proteins ◽  
Mass Spectrometry Analysis ◽  
Post Translational Modification ◽  
Related Factors ◽  
Spectrometry Analysis ◽  
Binding Domains ◽  
Family Protein ◽  
Related Proteins

Human GW182 family proteins have Argonaute (AGO)-binding domains in their N-terminal regions and silencing domains, which interact with RNA silencing-related proteins, in their C-terminal regions. Thus, they function as scaffold proteins between the AGO protein and RNA silencing-related proteins, such as carbon catabolite repressor4-negative on TATA (CCR4-NOT) or poly(A)-binding protein (PABP). Our mass spectrometry analysis and the phosphorylation data registered in PhosphoSitePlus, a post-translational modification database, suggested that the C-terminal region of a human GW182 family protein, TNRC6A, has at least four possible phosphorylation sites, which are located near the region interacting with the CCR4-NOT complex. Among them, two serine residues at amino acid positions 1332 and 1346 (S1332 and S1346) were certainly phosphorylated in human HeLa cells, but other two serine residues (S1616 and S1691) were not phosphorylated. Furthermore, it was revealed that the phosphorylation patterns of TNRC6A affect the interaction with the CCR4-NOT complex. When S1332 and S1346 were dephosphorylated, the interactions of TNRC6A with the CCR4-NOT complex were enhanced, and when S1616 and S1691 were phosphorylated, such interaction was suppressed. Thus, phosphorylation of TNRC6A was considered to regulate the interaction with RNA silencing-related factors that may affect RNA silencing activity.

Spectrophotometry Measurement of Polynuclear Aromatic Hydrocarbons in Hamilton Harbour Sediments

1991 ◽  
Vol 26 (1) ◽  
pp. 1-16 ◽  
Author(s):  
T.P. Murphy ◽  
H. Brouwer ◽  
M.E. Fox ◽  
E. Nagy
Keyword(s):  
Aromatic Hydrocarbons ◽  
Total Concentration ◽  
Coal Tar ◽  
Sediment Cores ◽  
Polynuclear Aromatic Hydrocarbons ◽  
Mass Spectrometry Analysis ◽  
Gas Chromatography Mass Spectrometry ◽  
Near Surface ◽  
Spectrometry Analysis ◽  
Hamilton Harbour

Abstract Eighty-one sediment cores were collected to determine the extent of coal tar contamination in a toxic area of Hamilton Harbour. Over 800 samples were analyzed by a UV spectrophotometric technique that was standardized with gas chromatography/mass spectrometry analysis. The coal tar distribution was variable. The highest concentrations were near the Stelco outfalls and the Hamilton-Wentworth combined sewer outfalls. The total concentration of the 16 polynuclear aromatic hydrocarbons (PAHs) in 48,300 m3 of near-surface sediments exceeded 200 µg/g.

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