A Dictyostelium SH2 adaptor protein required for correct DIF-1 signaling and pattern formation

Christopher Sugden, Susan Ross, Sarah J. Annesley, Christian Cole, Gareth Bloomfield, Alasdair Ivens, Jason Skelton, Paul R. Fisher, Geoffrey Barton, Jeffrey G. Williams

    Research output: Contribution to journalArticle

    1 Citation (Scopus)

    Abstract

    Dictyostelium is the only non-metazoan with functionally analyzed SH2 domains and studying them can give insights into their evolution and wider potential. LrrB has a novel domain configuration with leucine-rich repeat, 14-3-3 and SH2 protein-protein interaction modules. It is required for the correct expression of several specific genes in early development and here we characterize its role in later, multicellular development. During development in the light, slug formation in LrrB null (IrrB-) mutants is delayed relative to the parental strain, and the slugs are highly defective in phototaxis and thermotaxis. In the dark the mutant arrests development as an elongated mound, in a hitherto unreported process we term dark stalling. The developmental and phototaxis defects are cell autonomous and marker analysis shows that the pstO prestalk sub-region of the slug is aberrant in the IrrB- mutant. Expression profiling, by parallel micro-array and deep RNA sequence analyses, reveals many other alterations in prestalk-specific gene expression in IrrB- slugs, including reduced expression of the ecmB gene and elevated expression of ampA. During culmination ampA is ectopically expressed in the stalk, there is no expression of ampA and ecmB in the lower cup and the mutant fruiting bodies lack a basal disc. The basal disc cup derives from the pstB cells and this population is greatly reduced in the IrrB- mutant. This anatomical feature is a hallmark of mutants aberrant in signaling by DIF-1, the polyketide that induces prestalk and stalk cell differentiation. In a DIF-1 induction assay the IrrB- mutant is profoundly defective in ecmB activation but only marginally defective in ecmA induction. Thus the mutation partially uncouples these two inductive events. In early development LrrB interacts physically and functionally with CldA, another SH2 domain containing protein. However, the CldA null mutant does not phenocopy the IrrB- in its aberrant multicellular development or phototaxis defect, implying that the early and late functions of LrrB are affected in different ways. These observations, coupled with its domain structure, suggest that LrrB is an SH2 adaptor protein active in diverse developmental signaling pathways. (C) 2011 Elsevier Inc. All rights reserved.

    Original languageEnglish
    Pages (from-to)290-301
    Number of pages12
    JournalDevelopmental Biology
    Volume353
    Issue number2
    DOIs
    Publication statusPublished - 15 May 2011

    Keywords

    • Dictyostelium
    • SH2 domain
    • Adaptor protein
    • DIF-1
    • Prestalk
    • Basal disc
    • SLUG PHOTOTAXIS MUTANTS
    • ANTERIOR-LIKE CELLS
    • TRANSCRIPTION FACTOR
    • GENE-EXPRESSION
    • DOMAIN PROTEINS
    • BASAL DISC
    • RNA-SEQ
    • DISCOIDEUM
    • CULMINATION
    • GRB2

    Cite this

    Sugden, C., Ross, S., Annesley, S. J., Cole, C., Bloomfield, G., Ivens, A., ... Williams, J. G. (2011). A Dictyostelium SH2 adaptor protein required for correct DIF-1 signaling and pattern formation. Developmental Biology, 353(2), 290-301. https://doi.org/10.1016/j.ydbio.2011.03.003
    Sugden, Christopher ; Ross, Susan ; Annesley, Sarah J. ; Cole, Christian ; Bloomfield, Gareth ; Ivens, Alasdair ; Skelton, Jason ; Fisher, Paul R. ; Barton, Geoffrey ; Williams, Jeffrey G. / A Dictyostelium SH2 adaptor protein required for correct DIF-1 signaling and pattern formation. In: Developmental Biology. 2011 ; Vol. 353, No. 2. pp. 290-301.
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    abstract = "Dictyostelium is the only non-metazoan with functionally analyzed SH2 domains and studying them can give insights into their evolution and wider potential. LrrB has a novel domain configuration with leucine-rich repeat, 14-3-3 and SH2 protein-protein interaction modules. It is required for the correct expression of several specific genes in early development and here we characterize its role in later, multicellular development. During development in the light, slug formation in LrrB null (IrrB-) mutants is delayed relative to the parental strain, and the slugs are highly defective in phototaxis and thermotaxis. In the dark the mutant arrests development as an elongated mound, in a hitherto unreported process we term dark stalling. The developmental and phototaxis defects are cell autonomous and marker analysis shows that the pstO prestalk sub-region of the slug is aberrant in the IrrB- mutant. Expression profiling, by parallel micro-array and deep RNA sequence analyses, reveals many other alterations in prestalk-specific gene expression in IrrB- slugs, including reduced expression of the ecmB gene and elevated expression of ampA. During culmination ampA is ectopically expressed in the stalk, there is no expression of ampA and ecmB in the lower cup and the mutant fruiting bodies lack a basal disc. The basal disc cup derives from the pstB cells and this population is greatly reduced in the IrrB- mutant. This anatomical feature is a hallmark of mutants aberrant in signaling by DIF-1, the polyketide that induces prestalk and stalk cell differentiation. In a DIF-1 induction assay the IrrB- mutant is profoundly defective in ecmB activation but only marginally defective in ecmA induction. Thus the mutation partially uncouples these two inductive events. In early development LrrB interacts physically and functionally with CldA, another SH2 domain containing protein. However, the CldA null mutant does not phenocopy the IrrB- in its aberrant multicellular development or phototaxis defect, implying that the early and late functions of LrrB are affected in different ways. These observations, coupled with its domain structure, suggest that LrrB is an SH2 adaptor protein active in diverse developmental signaling pathways. (C) 2011 Elsevier Inc. All rights reserved.",
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    Sugden, C, Ross, S, Annesley, SJ, Cole, C, Bloomfield, G, Ivens, A, Skelton, J, Fisher, PR, Barton, G & Williams, JG 2011, 'A Dictyostelium SH2 adaptor protein required for correct DIF-1 signaling and pattern formation', Developmental Biology, vol. 353, no. 2, pp. 290-301. https://doi.org/10.1016/j.ydbio.2011.03.003

    A Dictyostelium SH2 adaptor protein required for correct DIF-1 signaling and pattern formation. / Sugden, Christopher; Ross, Susan; Annesley, Sarah J.; Cole, Christian; Bloomfield, Gareth; Ivens, Alasdair; Skelton, Jason; Fisher, Paul R.; Barton, Geoffrey; Williams, Jeffrey G.

    In: Developmental Biology, Vol. 353, No. 2, 15.05.2011, p. 290-301.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - A Dictyostelium SH2 adaptor protein required for correct DIF-1 signaling and pattern formation

    AU - Sugden, Christopher

    AU - Ross, Susan

    AU - Annesley, Sarah J.

    AU - Cole, Christian

    AU - Bloomfield, Gareth

    AU - Ivens, Alasdair

    AU - Skelton, Jason

    AU - Fisher, Paul R.

    AU - Barton, Geoffrey

    AU - Williams, Jeffrey G.

    PY - 2011/5/15

    Y1 - 2011/5/15

    N2 - Dictyostelium is the only non-metazoan with functionally analyzed SH2 domains and studying them can give insights into their evolution and wider potential. LrrB has a novel domain configuration with leucine-rich repeat, 14-3-3 and SH2 protein-protein interaction modules. It is required for the correct expression of several specific genes in early development and here we characterize its role in later, multicellular development. During development in the light, slug formation in LrrB null (IrrB-) mutants is delayed relative to the parental strain, and the slugs are highly defective in phototaxis and thermotaxis. In the dark the mutant arrests development as an elongated mound, in a hitherto unreported process we term dark stalling. The developmental and phototaxis defects are cell autonomous and marker analysis shows that the pstO prestalk sub-region of the slug is aberrant in the IrrB- mutant. Expression profiling, by parallel micro-array and deep RNA sequence analyses, reveals many other alterations in prestalk-specific gene expression in IrrB- slugs, including reduced expression of the ecmB gene and elevated expression of ampA. During culmination ampA is ectopically expressed in the stalk, there is no expression of ampA and ecmB in the lower cup and the mutant fruiting bodies lack a basal disc. The basal disc cup derives from the pstB cells and this population is greatly reduced in the IrrB- mutant. This anatomical feature is a hallmark of mutants aberrant in signaling by DIF-1, the polyketide that induces prestalk and stalk cell differentiation. In a DIF-1 induction assay the IrrB- mutant is profoundly defective in ecmB activation but only marginally defective in ecmA induction. Thus the mutation partially uncouples these two inductive events. In early development LrrB interacts physically and functionally with CldA, another SH2 domain containing protein. However, the CldA null mutant does not phenocopy the IrrB- in its aberrant multicellular development or phototaxis defect, implying that the early and late functions of LrrB are affected in different ways. These observations, coupled with its domain structure, suggest that LrrB is an SH2 adaptor protein active in diverse developmental signaling pathways. (C) 2011 Elsevier Inc. All rights reserved.

    AB - Dictyostelium is the only non-metazoan with functionally analyzed SH2 domains and studying them can give insights into their evolution and wider potential. LrrB has a novel domain configuration with leucine-rich repeat, 14-3-3 and SH2 protein-protein interaction modules. It is required for the correct expression of several specific genes in early development and here we characterize its role in later, multicellular development. During development in the light, slug formation in LrrB null (IrrB-) mutants is delayed relative to the parental strain, and the slugs are highly defective in phototaxis and thermotaxis. In the dark the mutant arrests development as an elongated mound, in a hitherto unreported process we term dark stalling. The developmental and phototaxis defects are cell autonomous and marker analysis shows that the pstO prestalk sub-region of the slug is aberrant in the IrrB- mutant. Expression profiling, by parallel micro-array and deep RNA sequence analyses, reveals many other alterations in prestalk-specific gene expression in IrrB- slugs, including reduced expression of the ecmB gene and elevated expression of ampA. During culmination ampA is ectopically expressed in the stalk, there is no expression of ampA and ecmB in the lower cup and the mutant fruiting bodies lack a basal disc. The basal disc cup derives from the pstB cells and this population is greatly reduced in the IrrB- mutant. This anatomical feature is a hallmark of mutants aberrant in signaling by DIF-1, the polyketide that induces prestalk and stalk cell differentiation. In a DIF-1 induction assay the IrrB- mutant is profoundly defective in ecmB activation but only marginally defective in ecmA induction. Thus the mutation partially uncouples these two inductive events. In early development LrrB interacts physically and functionally with CldA, another SH2 domain containing protein. However, the CldA null mutant does not phenocopy the IrrB- in its aberrant multicellular development or phototaxis defect, implying that the early and late functions of LrrB are affected in different ways. These observations, coupled with its domain structure, suggest that LrrB is an SH2 adaptor protein active in diverse developmental signaling pathways. (C) 2011 Elsevier Inc. All rights reserved.

    KW - Dictyostelium

    KW - SH2 domain

    KW - Adaptor protein

    KW - DIF-1

    KW - Prestalk

    KW - Basal disc

    KW - SLUG PHOTOTAXIS MUTANTS

    KW - ANTERIOR-LIKE CELLS

    KW - TRANSCRIPTION FACTOR

    KW - GENE-EXPRESSION

    KW - DOMAIN PROTEINS

    KW - BASAL DISC

    KW - RNA-SEQ

    KW - DISCOIDEUM

    KW - CULMINATION

    KW - GRB2

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    DO - 10.1016/j.ydbio.2011.03.003

    M3 - Article

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    EP - 301

    JO - Developmental Biology

    JF - Developmental Biology

    SN - 0012-1606

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    ER -