RCC1 isoforms differ in their affinity for chromatin, molecular interactions and regulation by phosphorylation

Fiona E. Hood, Paul R. Clarke

    Research output: Contribution to journalArticle

    23 Citations (Scopus)

    Abstract

    RCC1 is the guanine nucleotide exchange factor for Ran GTPase. Generation of Ran-GTP by RCC1 on chromatin provides a spatial signal that directs nucleocytoplasmic transport, mitotic spindle assembly and nuclear envelope formation. We show that RCC1 is expressed in human cells as at least three isoforms, named RCC1a, RCC1ß and RCC1?, which are expressed at different levels in specific tissues. The ß and ? isoforms contain short inserts in their N-terminal regions (NTRs) that are not present in RCC1a. This region mediates interaction with chromatin, binds importin a3 and/or importin ß, and contains regulatory phosphorylation sites. RCC1? is predominantly localised to the nucleus and mitotic chromosomes like RCC1a. However, compared to RCC1a, RCC1? has a greatly reduced interaction with an importin a3-ß and a stronger interaction with chromatin that is mediated by the extended NTR. RCC1? is also the isoform that is most highly phosphorylated at serine 11 in mitosis. Unlike RCC1a, RCC1? supports cell proliferation in tsBN2 cells more efficiently when serine 11 is mutated to non-phosphorylatable alanine. Phosphorylation of RCC1? therefore specifically controls its function during mitosis. These results show that human RCC1 isoforms have distinct chromatin binding properties, different molecular interactions, and are selectively regulated by phosphorylation, as determined by their different NTRs.

    Original languageEnglish
    Pages (from-to)3436-45
    Number of pages10
    JournalJournal of Cell Science
    Volume120
    Issue numberPt 19
    DOIs
    Publication statusPublished - 1 Oct 2007

    Fingerprint

    Karyopherins
    Chromatin
    Protein Isoforms
    Phosphorylation
    Mitosis
    Serine
    Guanine Nucleotide Exchange Factors
    Spindle Apparatus
    Cell Nucleus Active Transport
    GTP Phosphohydrolases
    Nuclear Envelope
    Guanosine Triphosphate
    Alanine
    Chromosomes
    Cell Proliferation

    Keywords

    • Amino Acid Sequence
    • Animals
    • Cell Cycle Proteins
    • Cell Line
    • Chromatin
    • Guanine Nucleotide Exchange Factors
    • Humans
    • Karyopherins
    • Molecular Sequence Data
    • Nuclear Proteins
    • Phosphorylation
    • Protein Binding
    • Protein Isoforms
    • Recombinant Fusion Proteins
    • Sequence Alignment
    • Tissue Distribution

    Cite this

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    abstract = "RCC1 is the guanine nucleotide exchange factor for Ran GTPase. Generation of Ran-GTP by RCC1 on chromatin provides a spatial signal that directs nucleocytoplasmic transport, mitotic spindle assembly and nuclear envelope formation. We show that RCC1 is expressed in human cells as at least three isoforms, named RCC1a, RCC1{\ss} and RCC1?, which are expressed at different levels in specific tissues. The {\ss} and ? isoforms contain short inserts in their N-terminal regions (NTRs) that are not present in RCC1a. This region mediates interaction with chromatin, binds importin a3 and/or importin {\ss}, and contains regulatory phosphorylation sites. RCC1? is predominantly localised to the nucleus and mitotic chromosomes like RCC1a. However, compared to RCC1a, RCC1? has a greatly reduced interaction with an importin a3-{\ss} and a stronger interaction with chromatin that is mediated by the extended NTR. RCC1? is also the isoform that is most highly phosphorylated at serine 11 in mitosis. Unlike RCC1a, RCC1? supports cell proliferation in tsBN2 cells more efficiently when serine 11 is mutated to non-phosphorylatable alanine. Phosphorylation of RCC1? therefore specifically controls its function during mitosis. These results show that human RCC1 isoforms have distinct chromatin binding properties, different molecular interactions, and are selectively regulated by phosphorylation, as determined by their different NTRs.",
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    author = "Hood, {Fiona E.} and Clarke, {Paul R.}",
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    language = "English",
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    RCC1 isoforms differ in their affinity for chromatin, molecular interactions and regulation by phosphorylation. / Hood, Fiona E.; Clarke, Paul R.

    In: Journal of Cell Science, Vol. 120, No. Pt 19, 01.10.2007, p. 3436-45.

    Research output: Contribution to journalArticle

    TY - JOUR

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    AU - Hood, Fiona E.

    AU - Clarke, Paul R.

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    N2 - RCC1 is the guanine nucleotide exchange factor for Ran GTPase. Generation of Ran-GTP by RCC1 on chromatin provides a spatial signal that directs nucleocytoplasmic transport, mitotic spindle assembly and nuclear envelope formation. We show that RCC1 is expressed in human cells as at least three isoforms, named RCC1a, RCC1ß and RCC1?, which are expressed at different levels in specific tissues. The ß and ? isoforms contain short inserts in their N-terminal regions (NTRs) that are not present in RCC1a. This region mediates interaction with chromatin, binds importin a3 and/or importin ß, and contains regulatory phosphorylation sites. RCC1? is predominantly localised to the nucleus and mitotic chromosomes like RCC1a. However, compared to RCC1a, RCC1? has a greatly reduced interaction with an importin a3-ß and a stronger interaction with chromatin that is mediated by the extended NTR. RCC1? is also the isoform that is most highly phosphorylated at serine 11 in mitosis. Unlike RCC1a, RCC1? supports cell proliferation in tsBN2 cells more efficiently when serine 11 is mutated to non-phosphorylatable alanine. Phosphorylation of RCC1? therefore specifically controls its function during mitosis. These results show that human RCC1 isoforms have distinct chromatin binding properties, different molecular interactions, and are selectively regulated by phosphorylation, as determined by their different NTRs.

    AB - RCC1 is the guanine nucleotide exchange factor for Ran GTPase. Generation of Ran-GTP by RCC1 on chromatin provides a spatial signal that directs nucleocytoplasmic transport, mitotic spindle assembly and nuclear envelope formation. We show that RCC1 is expressed in human cells as at least three isoforms, named RCC1a, RCC1ß and RCC1?, which are expressed at different levels in specific tissues. The ß and ? isoforms contain short inserts in their N-terminal regions (NTRs) that are not present in RCC1a. This region mediates interaction with chromatin, binds importin a3 and/or importin ß, and contains regulatory phosphorylation sites. RCC1? is predominantly localised to the nucleus and mitotic chromosomes like RCC1a. However, compared to RCC1a, RCC1? has a greatly reduced interaction with an importin a3-ß and a stronger interaction with chromatin that is mediated by the extended NTR. RCC1? is also the isoform that is most highly phosphorylated at serine 11 in mitosis. Unlike RCC1a, RCC1? supports cell proliferation in tsBN2 cells more efficiently when serine 11 is mutated to non-phosphorylatable alanine. Phosphorylation of RCC1? therefore specifically controls its function during mitosis. These results show that human RCC1 isoforms have distinct chromatin binding properties, different molecular interactions, and are selectively regulated by phosphorylation, as determined by their different NTRs.

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    KW - Karyopherins

    KW - Molecular Sequence Data

    KW - Nuclear Proteins

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    KW - Protein Binding

    KW - Protein Isoforms

    KW - Recombinant Fusion Proteins

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    KW - Tissue Distribution

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    SN - 0021-9533

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