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Germinal Center Kinases SmKIN3 and SmKIN24 Are Associated with the Sordaria macrospora Striatin-Interacting Phosphatase and Kinase (STRIPAK) Complex.

Frey S, Reschka EJ, Pöggeler S - PLoS ONE (2015)

Bottom Line: Here, we report on the presence and function of STRIPAK-associated kinases in ascomycetes.The physical interaction of the striatin homolog PRO11 with SmKIN3 and SmKIN24 were verified by yeast two-hybrid (Y2H) interaction studies and for SmKIN3 by co-Immunoprecipitation (co-IP).In vivo localization found that both kinases were present at the septa and deletion of both Smkin3 and Smkin24 led to abnormal septum distribution.

View Article: PubMed Central - PubMed

Affiliation: Institute of Microbiology and Genetics, Department of Genetics of Eukaryotic Microorganisms, Georg-August-University Göttingen, Göttingen, Germany.

ABSTRACT
The striatin-interacting phosphatase and kinase (STRIPAK) complex is composed of striatin, protein phosphatase PP2A and protein kinases that regulate development in animals and fungi. In the filamentous ascomycete Sordaria macrospora, it is required for fruiting-body development and cell fusion. Here, we report on the presence and function of STRIPAK-associated kinases in ascomycetes. Using the mammalian germinal center kinases (GCKs) MST4, STK24, STK25 and MINK1 as query, we identified the two putative homologs SmKIN3 and SmKIN24 in S. macrospora. A BLASTP search revealed that both kinases are conserved among filamentous ascomycetes. The physical interaction of the striatin homolog PRO11 with SmKIN3 and SmKIN24 were verified by yeast two-hybrid (Y2H) interaction studies and for SmKIN3 by co-Immunoprecipitation (co-IP). In vivo localization found that both kinases were present at the septa and deletion of both Smkin3 and Smkin24 led to abnormal septum distribution. While deletion of Smkin3 caused larger distances between adjacent septa and increased aerial hyphae, deletion of Smkin24 led to closer spacing of septa and to sterility. Although phenotypically distinct, both kinases appear to function independently because the double-knockout strain ΔSmkin3/ΔSmkin24 displayed the combined phenotypes of each single-deletion strain.

No MeSH data available.


Related in: MedlinePlus

Schematic representation of mammalian, D. melanogaster and fungal STRIPAK complex and STRIPAK-like complexes (A) and structure of the Hippo network in animals and the MEN/SIN as well as the RAM/MOR signaling pathway in fungi (B).(A) Core components of the STRIPAK and STRIPAK-like complex (striatin, Mob3, catalytic and scaffold subunit of the phosphatase PP2A, and STRIP proteins and the respective homologs) are depicted in green, putative homologs of the cortactin-binding protein CTTNBP2 are shown in red, putative homologs of SLMAP in light blue and putative SIKE/FGFR1OP2 in dark blue. STRIPAK GCKs and the associated CCM3 protein are indicated in violet. The arrangements of the subunits in the respective are deduced from [15, 20, 27–34]. (B) Components of the Hippo pathway in D. melanogaster and mammals are indicated in orange and green, respectively. Homologous components of the S. cerevisiae (Sc) MEN and RAM network and the corresponding S. pombe (Sp) SIN and MOR network are shown in the left and right boxes. Components from the filamentous fungus N. crassa (Nc) are shown in red according to [35]. The figure was modified from [23].
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pone.0139163.g001: Schematic representation of mammalian, D. melanogaster and fungal STRIPAK complex and STRIPAK-like complexes (A) and structure of the Hippo network in animals and the MEN/SIN as well as the RAM/MOR signaling pathway in fungi (B).(A) Core components of the STRIPAK and STRIPAK-like complex (striatin, Mob3, catalytic and scaffold subunit of the phosphatase PP2A, and STRIP proteins and the respective homologs) are depicted in green, putative homologs of the cortactin-binding protein CTTNBP2 are shown in red, putative homologs of SLMAP in light blue and putative SIKE/FGFR1OP2 in dark blue. STRIPAK GCKs and the associated CCM3 protein are indicated in violet. The arrangements of the subunits in the respective are deduced from [15, 20, 27–34]. (B) Components of the Hippo pathway in D. melanogaster and mammals are indicated in orange and green, respectively. Homologous components of the S. cerevisiae (Sc) MEN and RAM network and the corresponding S. pombe (Sp) SIN and MOR network are shown in the left and right boxes. Components from the filamentous fungus N. crassa (Nc) are shown in red according to [35]. The figure was modified from [23].

Mentions: One of these pro genes, namely pro11, encodes a homolog of the mammalian striatin protein family [8]. Mammalian striatins are multidomain proteins possessing a caveolin-binding domain, a coiled-coil domain, a calmodulin binding domain at the N-terminus and a C-terminal WD40 repeat region. They were initially identified as a novel group of phosphatase PP2A regulatory B”‘ subunits with roles in both signaling and trafficking [14]. Moreover, striatins serve as scaffolding units in the striatin-interacting phosphatase and kinase (STRIPAK) complex [15] (Fig 1A). In mammals, this recently identified protein complex consists of striatin as scaffold, the putative kinase activator monopolar spindle-one-binder 3 (MOB3), serine/threonine-phosphatase PP2A subunits A and C, cerebral cavernous malformation 3 (CCM3) protein, the striatin-interacting protein (STRIP)1 and STRIP2 and the germinal center kinases (GCK) MST4, STK24, STK25 and MINK1 [15–19]. STRIPAK sub-complexes and STRIPAK-like complexes have since been identified in other organisms. In mammals, the STRIPAK sub-complex mutually exclusively interacts with the sarcolemmal membrane-associated protein (SLMAP), the related coiled-coil proteins suppressor of IKKepsilon (SIKE) and fibroblast growth factor receptor oncogene partner 2 (FGFR1OP2) or the cortactin-binding protein CTTNBP2 [15] (Fig 1A). In Drosophila melanogaster and mammals proteomic approaches identified the STRIPAK-like as a negative regulator of the Hippo signaling [20, 21]. The Hippo pathways consists of a kinase cascade that regulates tissue and organ size in metazoans [22] (Fig 1B). Core components of this pathways are highly conserved in the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe as well as in filamentous fungi [23, 24]. In S. cerevisiae and fission yeast, one pathway is involved in coupling the cell cycle and cell septation and is termed the MEN (mitotic exit network) and SIN (septation initiation network), respectively [23, 25]. The second kinase pathway, which regulates morphology and polar growth is termed RAM (regulation of Ace2 and morphogenesis) in S. cerevisiae and MOR (morphogenesis of Orb6) in S. pombe [23, 26] (Fig 1B).


Germinal Center Kinases SmKIN3 and SmKIN24 Are Associated with the Sordaria macrospora Striatin-Interacting Phosphatase and Kinase (STRIPAK) Complex.

Frey S, Reschka EJ, Pöggeler S - PLoS ONE (2015)

Schematic representation of mammalian, D. melanogaster and fungal STRIPAK complex and STRIPAK-like complexes (A) and structure of the Hippo network in animals and the MEN/SIN as well as the RAM/MOR signaling pathway in fungi (B).(A) Core components of the STRIPAK and STRIPAK-like complex (striatin, Mob3, catalytic and scaffold subunit of the phosphatase PP2A, and STRIP proteins and the respective homologs) are depicted in green, putative homologs of the cortactin-binding protein CTTNBP2 are shown in red, putative homologs of SLMAP in light blue and putative SIKE/FGFR1OP2 in dark blue. STRIPAK GCKs and the associated CCM3 protein are indicated in violet. The arrangements of the subunits in the respective are deduced from [15, 20, 27–34]. (B) Components of the Hippo pathway in D. melanogaster and mammals are indicated in orange and green, respectively. Homologous components of the S. cerevisiae (Sc) MEN and RAM network and the corresponding S. pombe (Sp) SIN and MOR network are shown in the left and right boxes. Components from the filamentous fungus N. crassa (Nc) are shown in red according to [35]. The figure was modified from [23].
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4587736&req=5

pone.0139163.g001: Schematic representation of mammalian, D. melanogaster and fungal STRIPAK complex and STRIPAK-like complexes (A) and structure of the Hippo network in animals and the MEN/SIN as well as the RAM/MOR signaling pathway in fungi (B).(A) Core components of the STRIPAK and STRIPAK-like complex (striatin, Mob3, catalytic and scaffold subunit of the phosphatase PP2A, and STRIP proteins and the respective homologs) are depicted in green, putative homologs of the cortactin-binding protein CTTNBP2 are shown in red, putative homologs of SLMAP in light blue and putative SIKE/FGFR1OP2 in dark blue. STRIPAK GCKs and the associated CCM3 protein are indicated in violet. The arrangements of the subunits in the respective are deduced from [15, 20, 27–34]. (B) Components of the Hippo pathway in D. melanogaster and mammals are indicated in orange and green, respectively. Homologous components of the S. cerevisiae (Sc) MEN and RAM network and the corresponding S. pombe (Sp) SIN and MOR network are shown in the left and right boxes. Components from the filamentous fungus N. crassa (Nc) are shown in red according to [35]. The figure was modified from [23].
Mentions: One of these pro genes, namely pro11, encodes a homolog of the mammalian striatin protein family [8]. Mammalian striatins are multidomain proteins possessing a caveolin-binding domain, a coiled-coil domain, a calmodulin binding domain at the N-terminus and a C-terminal WD40 repeat region. They were initially identified as a novel group of phosphatase PP2A regulatory B”‘ subunits with roles in both signaling and trafficking [14]. Moreover, striatins serve as scaffolding units in the striatin-interacting phosphatase and kinase (STRIPAK) complex [15] (Fig 1A). In mammals, this recently identified protein complex consists of striatin as scaffold, the putative kinase activator monopolar spindle-one-binder 3 (MOB3), serine/threonine-phosphatase PP2A subunits A and C, cerebral cavernous malformation 3 (CCM3) protein, the striatin-interacting protein (STRIP)1 and STRIP2 and the germinal center kinases (GCK) MST4, STK24, STK25 and MINK1 [15–19]. STRIPAK sub-complexes and STRIPAK-like complexes have since been identified in other organisms. In mammals, the STRIPAK sub-complex mutually exclusively interacts with the sarcolemmal membrane-associated protein (SLMAP), the related coiled-coil proteins suppressor of IKKepsilon (SIKE) and fibroblast growth factor receptor oncogene partner 2 (FGFR1OP2) or the cortactin-binding protein CTTNBP2 [15] (Fig 1A). In Drosophila melanogaster and mammals proteomic approaches identified the STRIPAK-like as a negative regulator of the Hippo signaling [20, 21]. The Hippo pathways consists of a kinase cascade that regulates tissue and organ size in metazoans [22] (Fig 1B). Core components of this pathways are highly conserved in the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe as well as in filamentous fungi [23, 24]. In S. cerevisiae and fission yeast, one pathway is involved in coupling the cell cycle and cell septation and is termed the MEN (mitotic exit network) and SIN (septation initiation network), respectively [23, 25]. The second kinase pathway, which regulates morphology and polar growth is termed RAM (regulation of Ace2 and morphogenesis) in S. cerevisiae and MOR (morphogenesis of Orb6) in S. pombe [23, 26] (Fig 1B).

Bottom Line: Here, we report on the presence and function of STRIPAK-associated kinases in ascomycetes.The physical interaction of the striatin homolog PRO11 with SmKIN3 and SmKIN24 were verified by yeast two-hybrid (Y2H) interaction studies and for SmKIN3 by co-Immunoprecipitation (co-IP).In vivo localization found that both kinases were present at the septa and deletion of both Smkin3 and Smkin24 led to abnormal septum distribution.

View Article: PubMed Central - PubMed

Affiliation: Institute of Microbiology and Genetics, Department of Genetics of Eukaryotic Microorganisms, Georg-August-University Göttingen, Göttingen, Germany.

ABSTRACT
The striatin-interacting phosphatase and kinase (STRIPAK) complex is composed of striatin, protein phosphatase PP2A and protein kinases that regulate development in animals and fungi. In the filamentous ascomycete Sordaria macrospora, it is required for fruiting-body development and cell fusion. Here, we report on the presence and function of STRIPAK-associated kinases in ascomycetes. Using the mammalian germinal center kinases (GCKs) MST4, STK24, STK25 and MINK1 as query, we identified the two putative homologs SmKIN3 and SmKIN24 in S. macrospora. A BLASTP search revealed that both kinases are conserved among filamentous ascomycetes. The physical interaction of the striatin homolog PRO11 with SmKIN3 and SmKIN24 were verified by yeast two-hybrid (Y2H) interaction studies and for SmKIN3 by co-Immunoprecipitation (co-IP). In vivo localization found that both kinases were present at the septa and deletion of both Smkin3 and Smkin24 led to abnormal septum distribution. While deletion of Smkin3 caused larger distances between adjacent septa and increased aerial hyphae, deletion of Smkin24 led to closer spacing of septa and to sterility. Although phenotypically distinct, both kinases appear to function independently because the double-knockout strain ΔSmkin3/ΔSmkin24 displayed the combined phenotypes of each single-deletion strain.

No MeSH data available.


Related in: MedlinePlus