Experimental evidence of a new mechanism of activation of bacterial histidine kinases

Version 1.0
Experimental evidence of a new mechanism of activation of bacterial histidine kinases
Bortolotti, Ana; Almada, Juan Cruz; Porrini, Lucía; Albanesi, Daniela; Miguel, Virginia; Cybulski, Larisa, 2024, "Experimental evidence of a new mechanism of activation of bacterial histidine kinases", https://doi.org/10.57715/UNR/MPA7NB, RDA UNR, V1, UNF:6:MyNjHceGjMJ3qmUD9nYL3Q== [fileUNF]
Learn about Data Citation Standards.
    The dataset is too large to download. Please select the files you need from the files table.
Contact Owner Share
Dataset Metrics
313 Views
2 Downloads
0 Citations
Description

This data set shows a new mechanism of activation of histidine kinases that participate in signal transduction in bacteria. Experimental evidence was obtained after sequence homology analysis, generation of variant kinases, and their biochemical and structural studies. The techniques used included: construction of modified of the two kinases under study, DesK and EnvZ; transcriptional activation measurements by beta-galactosidase assays, bacterial membrane purification and subsequent, western blot, structural modeling, protein purification and autophosphorylation measurements with radioactively labeled ATP.

Value of the data:

Histidine kinases are widely distributed in bacteria and play a central role in two-component systems. These are fundamental for the functioning of cells since they are the key to activate and/or inhibit primordial mechanisms by detecting environmental signals and responding adaptively. The mechanism by which this is possible has been established and accepted. In particular, this set of data enables a new perspective on the activation of these systems and can be used to compare with other bacterial kinases already described.

Descripción en español:

Este conjunto de datos muestra un nuevo mecanismo de activación de histidin quinasas que participan de la transducción de señales en bacterias. La evidencia experimental fue obtenida tras un análisis de homología de secuencia, generación y estudios bioquímico y estructural de quinasas variantes. Las técnicas utilizadas incluyeron: construcción de variantes modificadas de las dos quinasas en estudio, DesK y EnvZ; medidas de activación transcripcional por medio de ensayos de beta-galactosidasa, purificación de membranas bacterianas y posterior, western blot, modelado estructural, purificación de proteínas y medidas de autofosforilación con ATP marcado radioactivamente.

Reutilización de los datos:

Las Histidin quinasas están ampliamente distribuidas en bacterias y cumplen un rol central en los sistemas de dos componentes. Estos son fundamentales para el funcionamiento de las células ya que son la llave para activar y/o inhibir mecanismos primordiales detectando señales ambientales y responder de manera adaptativa. El mecanismo por el cual esto es posible ha sido establecido y aceptado. En particular, este conjunto de datos posibilita una nueva perspectiva en la activación de estos sistemas y se podrán utilizar para comparar con otras quinasas bacterianas ya descriptas.

Subject Chemistry; Medicine, Health and Life Sciences
Keyword Histidine Kinase, Enzyme Activation, Signal Transduction, Allosteric Regulation, consciousness, receptor, Histidina Quinasa, Activación Enzimática, Transducción de Señal, Regulación Alostérica
Related Publication Larisa Cybulski; Ana Bortolotti; Juan Cruz Almada; Ariel Fernandéz. Interhelical hydrogen bonds in the membrane core drive activity of transmembrane kinases. Argentina. 2021. Congreso. XLIX Reunión Anual de la Sociedad Argentina de Biofísica. isbn: 978-987-27591-9-3
License/Data Use Agreement

CC BY 4.0

Filter by
1 to 10 of 10 Files
9_DESK_DESR_H335Y_complex model.pdb
application/vnd.palm - 27.1 MB
Published Jul 1, 2024
0 Downloads
MD5: 882c896df053c365b150957752e0cb99
Full-length DesK-DesR complex performed by protein-protein docking, using HDOCK server. The DesK His188 was mutated to tyrosine in silico
8_DESK_DESR_WT_complex model.pdb
application/vnd.palm - 23.0 MB
Published Jul 1, 2024
0 Downloads
MD5: 52c3017fbd2667898741e8de386229f6
Full-length DesK-DesR complex performed by protein-protein docking, using HDOCK server.
7_DESK_GLU_protein model.pdb
application/vnd.palm - 14.0 MB
Published Jul 1, 2024
0 Downloads
MD5: 26e5011338644080dc253017b9d892e2
DesK structure in the phosphotransfer state (PDB 7JJI), where the H188E mutation was modified to H188Q.
6_DESK_HIS_protein model.pdb
application/vnd.palm - 12.7 MB
Published Jul 1, 2024
0 Downloads
MD5: a640b7c9c7edebabcbdc6af3497c9dfb
DesK structure in the phosphotransfer state (PDB 7JJI), where the H188E mutation was reverted to its WT.
5_EnvZ_wt_and_variant_activities_Bgal.tab
Tabular Data - 3.7 KB
Published Jul 1, 2024
0 Downloads
24 Variables, 27 Observations UNF:6:ZPUlJAEn/sRUWeYCsfZZZQ==
β-galactosidase activity from a PompC-lacZ reporter fusion was measured at low osmolarity (Sucrose 0%) or high osmolarity (Sucrose 15%)
Preview "5_EnvZ_wt_and_variant_activities_Bgal.tab"
4_Autophosphorylation_curve_DesK_wt_and_variant.tab
Tabular Data - 435 B
Published Jul 1, 2024
0 Downloads
3 Variables, 24 Observations UNF:6:AChpgPX6NtVSFF17J9LNVg==
Phosphorylation kinetics of DesKC WT and H335Q. Eight samples taken in the initial 30 seconds were analyzed with radioactive ATP
Preview "4_Autophosphorylation_curve_DesK_wt_and_variant.tab"
3_Western_blot-DesK_wt_and_variant.jpeg
3_Western_blot-DesK_wt_and_variant.jpeg
JPEG Image - 36.0 KB
Published Jul 1, 2024
2 Downloads
MD5: 7241e1860d16485d148aecf1c85b091b
Expression and membrane integration levels of DesK variants. Western blots of the membrane fraction of Bacillus cells expressing each DesK variant (DesK WT, H335A, H335Y, H335Q, H188Q/H335Q, H188E and H188Q).
Preview "3_Western_blot-DesK_wt_and_variant.jpeg"
2_DesK_wt_and_variant-activities_Bgal.tab
Tabular Data - 458 B
Published Jul 1, 2024
0 Downloads
10 Variables, 10 Observations UNF:6:+4DfY4QqrGJfiBAbXHTnww==
Signal translation in DesK and its variants. Bacillus subtilis ∆desk cells expressing either DesK WT and its variants H335A, H335Y, H188E, H188Q, H335Q or H188Q/H335Q were grown at 37 °C to an OD550 of 0.3, and then divided in two flasks. One was maintained at 37 °C (red bars), and the other transferred to 25 ºC (blue bars). β-galactosidase activity was measured in Miller Units (MU).
Preview "2_DesK_wt_and_variant-activities_Bgal.tab"
1_Sequence_alignment_of_DesK.tab
Tabular Data - 229.8 KB
Published Jul 1, 2024
0 Downloads
224 Variables, 306 Observations UNF:6:JChUDRZ6/doYy9jW2t+zgQ==
Homologs of DesK in different organisms. Sequence alignment get from a BLASTP using DesKC as query and excluding other Bacilus from the data base by CLUSTALW
Preview "1_Sequence_alignment_of_DesK.tab"
10_DESK_DESR_H335Q_complex model.pdb
application/vnd.palm - 27.1 MB
Published Jul 1, 2024
0 Downloads
MD5: 8887b43113362d30efe65835ec76ad2a
Full-length DesK-DesR complex performed by protein-protein docking, using HDOCK server. The DesK His188 was mutated to glutamine in silico
Edit File

This file has already been deleted (or replaced) in the current version. It may not be edited.

Restrict Files

Restricting limits access to published files. You can add or edit Terms of Access for Restricted Files and allow people to Request Access to those files.

Enable access request
Edit Embargo

The selected file or files have already been published. Contact an administrator to change the embargo date or reason of the file or files.
Delete Files

The file will be deleted after you click on the Delete button.

Files will not be removed from previously published versions of the dataset.

Select File(s)

Please select one or more files.

Share Dataset

Share this dataset on your favorite social media networks.

Dataset Citations

Citations for this dataset are retrieved from Crossref via DataCite using Make Data Count standards. For more information about dataset metrics, please refer to the User Guide.

Sorry, no citations were found.
Restricted Files Selected

The selected file(s) may not be downloaded because you have not been granted access.

Download Options

The files selected are too large to download as a ZIP.

You can select individual files that are below the 100.0 MB download limit from the files table, or use the Data Access API for programmatic access to the files.

Select File(s)

Please select a file or files to be downloaded.

Restricted Files Selected

The restricted file(s) selected may not be downloaded because you have not been granted access.

Click Continue to download the files you have access to download.

Delete Dataset

Are you sure you want to delete this dataset and all of its files? You cannot undelete this dataset.

Delete Draft Version

Are you sure you want to delete this draft version? Files will be reverted to the most recently published version. You cannot undelete this draft.

Unpublished Dataset Private URL

Private URL can only be used with unpublished versions of datasets.

Unpublished Dataset Private URL

Are you sure you want to disable the Private URL? If you have shared the Private URL with others they will no longer be able to use it to access your unpublished dataset.

Delete Files

The file(s) will be deleted after you click on the Delete button.

Files will not be removed from previously published versions of the dataset.

Compute

This dataset contains restricted files you may not compute on because you have not been granted access.

Deaccession Dataset

Are you sure you want to deaccession? The selected version(s) will no longer be viewable by the public.

Deaccession Dataset

Are you sure you want to deaccession this dataset? It will no longer be viewable by the public.

Version Differences Details

Please select two versions to view the differences.

Version Differences Details
 
Version:
Last Updated:
Version:
Last Updated:
Select File(s)

Please select a file or files for access request.

Select File(s)

Embargoed files cannot be accessed. Please select an unembargoed file or files for your access request.

Edit Tags

Select existing file tags or create new tags to describe your files. Each file can have more than one tag.

Request Access

  You need to Sign Up or Log In to request access.

Dataset Terms

This dataset is made available under the following terms. Please confirm and/or complete the information needed below in order to continue.

Our Community Norms as well as good scientific practices expect that proper credit is given via citation. Please use the data citation shown on the dataset page.

CC BY 4.0

Preview Guestbook

Upon downloading files the guestbook asks for the following information.

Account Information

Package File Download

Use the Download URL in a Wget command or a download manager to download this package file. Download via web browser is not recommended. User Guide - Downloading a Dataverse Package via URL

https://dataverse.unr.edu.ar/api/access/datafile/

Request Access

Please confirm and/or complete the information needed below in order to request access to files in this dataset.

Compute Batch
Clear Batch
Dataset Dataset Persistent ID Change Compute Batch
Compute Batch
Submit for Review

You will not be able to make changes to this dataset while it is in review.

Publish Dataset

Are you sure you want to republish this dataset?

Select if this is a minor or major version update.

CC BY 4.0

Creative Commons Attribution 4.0 International License.
Publish Dataset

This dataset cannot be published until Facultad de Ciencias Bioquímicas y Farmacéuticas is published by its administrator.

Publish Dataset

This dataset cannot be published until Facultad de Ciencias Bioquímicas y Farmacéuticas and RDA UNR are published.

Return to Author

Return this dataset to contributor for modification.