Related Experiment Video

Updated: Apr 21, 2026

Studying RNA Interactors of Protein Kinase RNA-Activated during the Mammalian Cell Cycle
10:05

Studying RNA Interactors of Protein Kinase RNA-Activated during the Mammalian Cell Cycle

Published on: March 5, 2019

7.0K

Phosphorylation-dependent interaction between a serine/threonine kinase PknA and a putative cell division protein

Jae Jin Lee1, Choon-Ming Kan, Jung Hun Lee

  • 1National Leading Research Laboratory, Department of Biological Sciences, Myongji University, Yongin, Gyeonggido, Republic of Korea.

The New Microbiologica
|November 12, 2014
PubMed

Insights

Mycobacterium tuberculosis serine/threonine protein kinase A (PknA) phosphorylates cell division protein Wag31. This interaction

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • Mycobacterium tuberculosis possesses eleven serine/threonine protein kinases (STPKs).
  • PknA is a crucial STPK regulating cell shape and division through protein phosphorylation.
  • Wag31, a cell division protein, was identified as a novel substrate for PknA.

Purpose of the Study:

  • To investigate the interaction between PknA and Wag31.
  • To determine if the phosphorylation state of Wag31 affects its interaction with PknA.
  • To explore potential novel drug targets for tuberculosis.

Main Methods:

  • Cloning and expression of PknA, wild-type Wag31 (Wag31WT), phosphoablative Wag31T73A, and phosphomimetic Wag31T73E.
  • Purification of PknA and Wag31 variants.
  • Far-western blot analysis to assess protein-protein interactions.

Main Results:

  • Far-western analysis confirmed a direct interaction between PknA and Wag31.
  • The interaction between PknA and Wag31 is dependent on the phosphorylation state of Wag31.
  • Specific phosphorylation states of Wag31 modulate its binding to PknA.

Conclusions:

  • The phosphorylation-dependent interaction between PknA and Wag31 is critical for M. tuberculosis growth regulation.
  • This interaction represents a potential novel target for anti-tuberculosis drug development.
  • Understanding this pathway could lead to new therapeutic strategies against tuberculosis.

More Related Videos

Assaying Protein Kinase Activity with Radiolabeled ATP08:05

Assaying Protein Kinase Activity with Radiolabeled ATP

Published on: May 26, 2017

19.6K
Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay12:26

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay

Published on: May 3, 2018

19.5K

Related Concept Videos

PI3K/mTOR/AKT Signaling Pathway01:22

PI3K/mTOR/AKT Signaling Pathway

The mammalian target of rapamycin  (mTOR) is a serine/threonine kinase that regulates growth, proliferation, and cell survival in response to hormones, growth factors, or nutrient availability. This kinase exists in two structurally and functionally distinct forms: mTOR complex 1  (mTORC1) and mTOR complex 2  (mTORC2). The first form (mTORC1) is composed of a rapamycin-sensitive Raptor and proline-rich Akt substrate, PRAS40. In contrast,  mTORC2 consists of a...
6.5K
MAPK Signaling Cascades01:07

MAPK Signaling Cascades

Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...
9.5K
The JAK-STAT Signaling Pathway01:20

The JAK-STAT Signaling Pathway

Several cytokine receptors have tightly bound Janus kinase or JAK proteins attached at their cytosolic tail. Small signaling molecules such as cytokines, growth hormones, or prolactins bind to the cytokine receptors and initiate their dimerization. The dimerization brings the cytosolic JAKs together that trans-phosphorylate and activates each other. The activated JAKs now phosphorylate cytosolic tails of the cytokine receptors, which serve as binding sites for adaptor proteins such as  SH2...
14.3K
Interactions Between Signaling Pathways01:19

Interactions Between Signaling Pathways

Signaling cascades usually lack linearity. Multiple pathways interact and regulate one another, allowing cells to integrate and respond to diverse environmental stimuli.
Convergence and divergence, and cross-talk between signaling pathways
Two distinct signaling pathways can converge on a single functional unit, which may either be a single protein or a complex of proteins. The response is either functionally distinct or synergistic between the two pathways but different from the response...
8.0K
cAMP-dependent Protein Kinase Pathways01:25

cAMP-dependent Protein Kinase Pathways

Cyclic Adenosine Monophosphate (cAMP) is an essential second messenger that activates protein kinase A (PKA) and regulates various biological processes. A single epinephrine molecule binds to GPCR and activates several heterotrimeric G proteins, each stimulating multiple adenylyl cyclase, amplifying the signal, and synthesizing large numbers of cAMP molecules. Small changes in cAMP concentration affect PKA activity. The binding of four cAMP molecules induces a conformational change in PKA,...
9.5K
mTOR Signaling and Cancer Progression03:03

mTOR Signaling and Cancer Progression

The mammalian target of rapamycin or mTOR protein was discovered in 1994 due to its direct interaction with rapamycin. The protein gets its name from a yeast homolog called TOR. The mTOR protein complex in mammalian cells plays a major role in balancing anabolic processes such as the synthesis of proteins, lipids, and nucleotides and catabolic processes, such as autophagy in response to environmental cues, such as availability of nutrients and growth factors.
The mTOR pathway or the...
5.2K