Protein interactome analysis of 12 mitogen-activated protein kinase kinase kinase in rice using a yeast two-hybrid system

PROTEOMICS ◽  
2014 ◽  
Vol 14 (1) ◽  
pp. 105-115 ◽  
Author(s):  
Raksha Singh ◽  
Jae-Eun Lee ◽  
Sarmina Dangol ◽  
Jihyun Choi ◽  
Ran Hee Yoo ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Hybrid System ◽  
Mitogen Activated Protein Kinase ◽  
Yeast Two Hybrid ◽  
Yeast Two Hybrid System ◽  
Protein Interactome ◽  
Mitogen Activated Protein ◽  
Two Hybrid ◽  
Protein Kinase Kinase

scholarly journals Rice Mitogen-Activated Protein Kinase Interactome Analysis Using the Yeast Two-Hybrid System

2012 ◽  
Vol 160 (1) ◽  
pp. 477-487 ◽  
Author(s):  
Raksha Singh ◽  
Mi-Ok Lee ◽  
Jae-Eun Lee ◽  
Jihyun Choi ◽  
Ji Hun Park ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Hybrid System ◽  
Mitogen Activated Protein Kinase ◽  
Yeast Two Hybrid ◽  
Yeast Two Hybrid System ◽  
Mitogen Activated Protein ◽  
Two Hybrid

scholarly journals PICK1: a perinuclear binding protein and substrate for protein kinase C isolated by the yeast two-hybrid system.

1995 ◽  
Vol 128 (3) ◽  
pp. 263-271 ◽  
Author(s):  
J Staudinger ◽  
J Zhou ◽  
R Burgess ◽  
S J Elledge ◽  
E N Olson
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Hybrid System ◽  
Catalytic Domain ◽  
Yeast Two Hybrid ◽  
Kinase C ◽  
Yeast Two Hybrid System ◽  
Wide Range ◽  
Two Hybrid

Protein kinase C (PKC) plays a central role in the control of proliferation and differentiation of a wide range of cell types by mediating the signal transduction response to hormones and growth factors. Upon activation by diacylglycerol, PKC translocates to different subcellular sites where it phosphorylates numerous proteins, most of which are unidentified. We used the yeast two-hybrid system to identify proteins that interact with activated PKC alpha. Using the catalytic region of PKC fused to the DNA binding domain of yeast GAL4 as "bait" to screen a mouse T cell cDNA library in which cDNA was fused to the GAL4 activation domain, we cloned several novel proteins that interact with C-kinase (PICKs). One of these proteins, designated PICK1, interacts specifically with the catalytic domain of PKC and is an efficient substrate for phosphorylation by PKC in vitro and in vivo. PICK1 is localized to the perinuclear region and is phosphorylated in response to PKC activation. PICK1 and other PICKs may play important roles in mediating the actions of PKC.

scholarly journals Ser-3 is important for regulating Mos interaction with and stimulation of mitogen-activated protein kinase kinase.

1995 ◽  
Vol 15 (9) ◽  
pp. 4727-4734 ◽  
Author(s):  
M Chen ◽  
J A Cooper
Keyword(s):  
Protein Kinase ◽  
Catalytic Domain ◽  
Mitogen Activated Protein Kinase ◽  
Early Embryos ◽  
Mitogen Activated Protein ◽  
Hybrid Test ◽  
Two Hybrid ◽  
Autophosphorylation Site ◽  
Protein Kinase Kinase

Mos is a germ cell-specific serine/threonine protein kinase that activates mitogen-activated protein kinase (MAPK) through MAPK kinase (MKK). In Xenopus oocytes, Mos synthesis is required for progesterone-induced activation of MAPK and maturation promoting factor. Injection of Mos or active MAPK causes mitotic arrest in early embryos, suggesting that Mos also acts via MKK and MAPK to induce the arrest of unfertilized eggs in metaphase of meiosis II. We have investigated whether Mos activity is regulated by phosphorylation. Previous studies have identified Ser-3 as the principal autophosphorylation site. We show that Mos interacts with the catalytic domain of MKK in a Saccharomyces cerevisiae two-hybrid test. Acidic substitutions of the sites phosphorylated by Mos in MKK reduce the interaction, implying that the complex may dissociate after phosphorylation of MKK by Mos. Furthermore, the Mos-MKK interaction requires Mos kinase activity, suggesting that Mos autophosphorylation may be involved in the interaction. Substitution of Ser-3 of Mos with Ala reduces the interaction with MKK and also reduces both the activation of MKK by Mos in vitro and cleavage arrest induced by Mos fusion protein in Xenopus embryos. By contrast, substitution of Ser-3 by Glu, an acidic amino acid that mimics phosphoserine, fosters the Mos interaction with MKK and permits activation of MKK in vitro and Mos-induced cleavage arrest. Moreover, the Glu-3 substitution increases the interaction of a kinase-inactive Mos mutant with MKK. Taken together, these results suggest that an important step in Mos activation involves the phosphorylation at Ser-3, which promotes Mos interaction with and activation of MKK.

scholarly journals Comprehensive analysis of mitogen-activated protein kinase cascades in chrysanthemum

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5037 ◽  
Author(s):  
Aiping Song ◽  
Yueheng Hu ◽  
Lian Ding ◽  
Xue Zhang ◽  
Peiling Li ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Expression Patterns ◽  
Defense Responses ◽  
Mitogen Activated Protein Kinase ◽  
Pcr Analysis ◽  
Yeast Two Hybrid ◽  
Qrt Pcr ◽  
Mapk Cascades ◽  
Mitogen Activated Protein ◽  
Two Hybrid

Background Mitogen-activated protein kinase (MAPK) cascades, an important type of pathway in eukaryotic signaling networks, play a key role in plant defense responses, growth and development. Methods Phylogenetic analysis and conserved motif analysis of the MKK and MPK families in Arabidopsis thaliana, Helianthus annuus and Chrysanthemum morifolium classified MKK genes and MPK genes. qRT-PCR was used for the expression patterns of CmMPK and CmMKK genes, and yeast two-hybrid assay was applied to clear the interaction between CmMPKs and CmMKKs. Results We characterized six MKK genes and 11 MPK genes in chrysanthemum based on transcriptomic sequences and classified these genes into four groups. qRT-PCR analysis demonstrated that CmMKKs and CmMPKs exhibited various expression patterns in different organs of chrysanthemum and in response to abiotic stresses and phytohormone treatments. Furthermore, a yeast two-hybrid assay was applied to analyze the interaction between CmMKKs and CmMPKs and reveal the MAPK cascades in chrysanthemum. Discussion Our data led us to propose that CmMKK4-CmMPK13 and CmMKK2-CmMPK4 may be involved in regulating salt resistance and in the relationship between CmMKK9 and CmMPK6 and temperature stress.

Analysis of interactions between the subunits of protein kinase CK2

1996 ◽  
Vol 74 (4) ◽  
pp. 541-547 ◽  
Author(s):  
David W. Litchfield ◽  
Elzbieta Slominski ◽  
Shawn Lewenza ◽  
Michael Narvey ◽  
Denis G. Bosc ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Hybrid System ◽  
Protein Kinase Ck2 ◽  
Casein Kinase Ii ◽  
Casein Kinase ◽  
Yeast Two Hybrid ◽  
Yeast Two Hybrid System ◽  
Kinase Ck2 ◽  
Α Subunits ◽  
Two Hybrid

Protein kinase CK2, which was formerly known as casein kinase II, is a highly conserved protein serine/threonine kinase implicated in the control of cell proliferation through its phosphorylation of regulatory nuclear proteins. The enzyme consists of catalytic (α and (or) α′) subunits and β subunits that modulate the activity of the catalytic subunits. These subunits are arranged in homotetrameric (i.e., α2β2 or α′2β2) or heterotetrameric (i.e., αα′β2) complexes. We previously demonstrated using the yeast two-hybrid system that α (or α′) subunits can interact with β subunits but not other α (or α′) subunits. By comparison, β subunits can interact with α (or α′) and with β subunits, suggesting that the protein kinase CK2 holoenzyme forms because of the ability of p subunits to dimerize, bringing two heterodimers (αβ or α′β) into a tetrameric complex. In the present study, we used the yeast two-hybrid system to examine the domains of interactions between the α and β subunits of protein kinase CK2. These studies indicate that the ability of β to interact with α resides within the carboxy-terminal domain of β. By comparison, our studies suggest that individual domains of α are not sufficient for interactions with β.Key words: protein kinase CK2, casein kinase II, yeast two-hybrid system, subunit interaction, signal transduction.

scholarly journals JSAP1, a Novel Jun N-Terminal Protein Kinase (JNK)-Binding Protein That Functions as a Scaffold Factor in the JNK Signaling Pathway

1999 ◽  
Vol 19 (11) ◽  
pp. 7539-7548 ◽  
Author(s):  
Michihiko Ito ◽  
Katsuji Yoshioka ◽  
Mizuho Akechi ◽  
Shinya Yamashita ◽  
Nobuhiko Takamatsu ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Mitogen Activated Protein Kinase ◽  
Yeast Two Hybrid ◽  
Terminal Protein ◽  
Jnk Signaling ◽  
Mapk Cascades ◽  
Mitogen Activated Protein ◽  
Jnk Signaling Pathway ◽  
Two Hybrid

ABSTRACT The major components of the mitogen-activated protein kinase (MAPK) cascades are MAPK, MAPK kinase (MAPKK), and MAPKK kinase (MAPKKK). Recent rapid progress in identifying members of MAPK cascades suggests that a number of such signaling pathways exist in cells. To date, however, how the specificity and efficiency of the MAPK cascades is maintained is poorly understood. Here, we have identified a novel mouse protein, termed Jun N-terminal protein kinase (JNK)/stress-activated protein kinase-associated protein 1 (JSAP1), by a yeast two-hybrid screen, using JNK3 MAPK as the bait. Of the mammalian MAPKs tested (JNK1, JNK2, JNK3, ERK2, and p38α), JSAP1 preferentially coprecipitated with the JNKs in cotransfected COS-7 cells. JNK3 showed a higher binding affinity for JSAP1, compared with JNK1 and JNK2. In similar cotransfection studies, JSAP1 also interacted with SEK1 MAPKK and MEKK1 MAPKKK, which are involved in the JNK cascades. The regions of JSAP1 that bound JNK, SEK1, and MEKK1 were distinct from one another. JNK and MEKK1 also bound JSAP1 in vitro, suggesting that these interactions are direct. In contrast, only the activated form of SEK1 associated with JSAP1 in cotransfected COS-7 cells. The unstimulated SEK1 bound to MEKK1; thus, SEK1 might indirectly associate with JSAP1 through MEKK1. Although JSAP1 coprecipitated with MEK1 MAPKK and Raf-1 MAPKKK, and not MKK6 or MKK7 MAPKK, in cotransfected COS-7 cells, MEK1 and Raf-1 do not interfere with the binding of SEK1 and MEKK1 to JSAP1, respectively. Overexpression of full-length JSAP1 in COS-7 cells led to a considerable enhancement of JNK3 activation, and modest enhancement of JNK1 and JNK2 activation, by the MEKK1-SEK1 pathway. Deletion of the JNK- or MEKK1-binding regions resulted in a significant reduction in the enhancement of the JNK3 activation in COS-7 cells. These results suggest that JSAP1 functions as a scaffold protein in the JNK3 cascade. We also discuss a scaffolding role for JSAP1 in the JNK1 and JNK2 cascades.

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