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      Cell cycle-dependent expression and subcellular localization of fructose 1,6-bisphosphatase

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          Abstract

          Recently a gluconeogenic enzyme was discovered—fructose 1,6-bisphosphatase (FBPase)—that localizes in the nucleus of a proliferating cell, but its physiological role in this compartment remains unclear. Here, we demonstrate the link between nuclear localization of FBPase and the cell cycle progression. Results of our studies indicate that in human and mouse squamous cell lung cancer, as well as in the HL-1 cardiomyocytes, FBPase nuclear localization correlates with nuclear localization of S and G2 phase cyclins. Additionally, activity and expression of the enzyme depends on cell cycle stages. Identification of FBPase interacting partners with mass spectrometry reveals a set of nuclear proteins involved in cell cycle regulation, mRNA processing and in stabilization of genomic DNA structure. To our knowledge, this is the first experimental evidence that muscle FBPase is involved in cell cycle events.

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          The online version of this article (doi:10.1007/s00418-011-0884-1) contains supplementary material, which is available to authorized users.

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          Most cited references33

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          HL-1 cells: a cardiac muscle cell line that contracts and retains phenotypic characteristics of the adult cardiomyocyte.

          W C Claycomb, N. Lanson, B Stallworth … (1998)
          We have derived a cardiac muscle cell line, designated HL-1, from the AT-1 mouse atrial cardiomyocyte tumor lineage. HL-1 cells can be serially passaged, yet they maintain the ability to contract and retain differentiated cardiac morphological, biochemical, and electrophysiological properties. Ultrastructural characteristics typical of embryonic atrial cardiac muscle cells were found consistently in the cultured HL-1 cells. Reverse transcriptase-PCR-based analyses confirmed a pattern of gene expression similar to that of adult atrial myocytes, including expression of alpha-cardiac myosin heavy chain, alpha-cardiac actin, and connexin43. They also express the gene for atrial natriuretic factor. Immunohistochemical staining of the HL-1 cells indicated that the distribution of the cardiac-specific markers desmin, sarcomeric myosin, and atrial natriuretic factor was similar to that of cultured atrial cardiomyocytes. A delayed rectifier potassium current (IKr) was the most prominent outward current in HL-1 cells. The activating currents displayed inward rectification and deactivating current tails were voltage-dependent, saturated at >+20 mV, and were highly sensitive to dofetilide (IC50 of 46.9 nM). Specific binding of [3H]dofetilide was saturable and fit a one-site binding isotherm with a Kd of 140 +/- 60 nM and a Bmax of 118 fmol per 10(5) cells. HL-1 cells represent a cardiac myocyte cell line that can be repeatedly passaged and yet maintain a cardiac-specific phenotype.
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            Overexpression of cyclin B1 in early-stage non-small cell lung cancer and its clinical implication.

            Kwang Hong, Hassan K. Hassan, L. Mao … (2000)
            Cyclin B1 is a key molecule for G2-M-phase transition during the cell cycle and is overexpressed in various tumor types. However, the expression status of cyclin B1 in lung cancer and its clinical significance remain unknown. We used immunohistochemistry studies to examine the expression of cyclin B1 in 77 non-small cell lung cancer specimens from patients with histological stage I disease. All of the patients underwent curative surgical treatment. The median length of follow-up care is 8.2 years. High-level cyclin B1 expression (a cyclin B1 labeling index > or =15%) was observed in 17 of the 77 (22%) tumors. Patients whose tumors expressed a high level of cyclin B1 had a significantly shorter survival time than patients whose tumors expressed a low level of cyclin B1 (P = 0.02, log-rank test). Interestingly, overexpression of cyclin B1 was more frequently observed in tumors with squamous cell histology than in tumors with other histological cell types (P = 0.01, Fisher's exact test). A subgroup analysis revealed that cyclin B1 overexpression seems to be an adverse prognostic factor only in patients with squamous cell carcinoma (SCC) of the lung (P = 0.02, log-rank test). Our data indicate that cyclin B1 may be dysregulated in non-small cell lung cancer, particularly in the SCC subtype, and that a high level of cyclin B1 expression may be a prognostic marker for patients with early-stage SCC of the lung.
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              Proteomic analysis of interchromatin granule clusters.

              Noriko Saitoh, Chris Spahr, Scott D. Patterson … (2004)
              A variety of proteins involved in gene expression have been localized within mammalian cell nuclei in a speckled distribution that predominantly corresponds to interchromatin granule clusters (IGCs). We have applied a mass spectrometry strategy to identify the protein composition of this nuclear organelle purified from mouse liver nuclei. Using this approach, we have identified 146 proteins, many of which had already been shown to be localized to IGCs, or their functions are common to other already identified IGC proteins. In addition, we identified 32 proteins for which only sequence information is available and thus these represent novel IGC protein candidates. We find that 54% of the identified IGC proteins have known functions in pre-mRNA splicing. In combination with proteins involved in other steps of pre-mRNA processing, 81% of the identified IGC proteins are associated with RNA metabolism. In addition, proteins involved in transcription, as well as several other cellular functions, have been identified in the IGC fraction. However, the predominance of pre-mRNA processing factors supports the proposed role of IGCs as assembly, modification, and/or storage sites for proteins involved in pre-mRNA processing.
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                Author and article information

                Contributors
                : +48-71-3759213 , +48-71-3759213 , drakus@biol.uni.wroc.pl
                Journal
                Journal ID (nlm-ta): Histochem Cell Biol
                Title: Histochemistry and Cell Biology
                Publisher: Springer-Verlag (Berlin/Heidelberg )
                ISSN (Print): 0948-6143
                ISSN (Electronic): 1432-119X
                Publication date (Electronic): 6 November 2011
                Publication date PMC-release: 6 November 2011
                Publication date (Print): January 2012
                Volume: 137
                Issue: 1
                Pages: 121-136
                Affiliations
                [1 ]Department of Animal Molecular Physiology, Institute of Zoology, Wroclaw University, Cybulskiego 30, 50-205 Wrocław, Poland
                [2 ]Department of Thoracic Surgery, Lower Silesian Pulmonary Center, Grabiszynska 105, 50-439 Wrocław, Poland
                Article
                Publisher ID: 884
                DOI: 10.1007/s00418-011-0884-1
                PMC ID: 3249204
                PubMed ID: 22057438
                SO-VID: 89a6e6a6-b90f-47de-84a2-70a3d422e04f
                Copyright © © The Author(s) 2011
                History
                Date accepted : 26 October 2011
                Categories
                Subject: Original Paper
                Custom metadata
                issue-copyright-statement © Springer-Verlag 2012

                ScienceOpen disciplines: Cell biology
                Keywords: kln-205,cyclins,hl-1,fbpase,squamous cell lung cancer
                Data availability:
                ScienceOpen disciplines: Cell biology
                Keywords: kln-205, cyclins, hl-1, fbpase, squamous cell lung cancer

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