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Differential dynamics of splicing factor SC35 during the cell cycle

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Abstract

Pre-mRNA splicing factors are enriched in nuclear domains termed interchromatin granule clusters or nuclear speckles. During mitosis, nuclear speckles are disassembled by metaphase and reassembled in telophase in structures termed mitotic interchromatin granules (MIGs). We analysed the dynamics of the splicing factor SC35 in interphase and mitotic cells. In HeLa cells expressing green fluorescent protein (GFP)-SC35, this was localized in speckles during interphase and dispersed in metaphase. In telophase, GFP-SC35 was highly enriched within telophase nuclei and also detected in MIGs. Fluorescence recovery after photobleaching (FRAP) experiments revealed that the mobility of GFP-SC35 was distinct in different mitotic compartments. Interestingly, the mobility of GFP-SC35 was 3-fold higher in the cytoplasm of metaphase cells compared with interphase speckles, the nucleoplasm or MIGs. Treatment of cells with inhibitors of cyclin-dependent kinases (cdks) caused changes in the organization of nuclear compartments such as nuclear speckles and nucleoli, with corresponding changes in the mobility of GFP-SC35 and GFP-fibrillarin. Our results suggest that the dynamics of SC35 are significantly influenced by the organization of the compartment in which it is localized during the cell cycle.

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Abbreviations

ASF:

alternate splicing factor

cdk:

cyclin-dependent kinase

Clk:

Cdc2-like kinase

DAPI:

4′,6-diamidino-2-phenylindole

FBS:

foetal bovine serum

FITC:

fluorescein isothiocyanate

FRAP:

fluorescence recovery after photobleaching

GFP:

green fluorescent protein

GST:

glutathione S-transferase

IGC:

interchromatin granule cluster

IPTG:

isopropyl-β-D-thiogalactopyranoside

mAb:

monoclonal antibody

MIG:

mitotic interchromatin granule

NAP:

NOR-associated patch

NOR:

nucleolar organizing region

PBS:

phosphate-buffered saline

PVDF:

polyvinylidene fluoride

RFI:

relative fluorescence intensity

RS:

serine-rich

SDS-PAGE:

sodium dodecyl sulphate-polyacrylamide gel electrophoresis

snRNP:

small nuclear ribonucleoprotein

snoRNA:

small nucleolar RNA

SRPK 1:

serine/arginine-rich protein kinase 1

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Authors and Affiliations

  1. Centre for Cellular and Molecular Biology, Hyderabad, 500 007, India

    Kaushlendra Tripathi & Veena K. Parnaik

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  1. Kaushlendra Tripathi
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  2. Veena K. Parnaik
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Correspondence to Veena K. Parnaik.

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Tripathi, K., Parnaik, V.K. Differential dynamics of splicing factor SC35 during the cell cycle. J Biosci 33, 345–354 (2008). https://doi.org/10.1007/s12038-008-0054-3

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