Biomedical subjects

Heike Laman

Publications and source records attributed to Heike Laman.

6 recordsLinked to original sources

PubMed2007

Small-molecule mimics of an alpha-helix for efficient transport of proteins into cells.

We designed and synthesized small-molecule mimics of an alpha-helical peptide protein transduction domain (PTD). These small-molecule carriers, which we termed SMoCs, are easily coupled to biomolecules, and efficiently deliver dye molecules and recombinant proteins into a variety of cell types. We designed the SMoCs using molecular modeling techniques. As an example of a protein cargo, we applied this new technology to the internalization of the DNA replication licensing repressor geminin, in vitro, providing evidence that extracellularly delivered SMoC-geminin can have an antiproliferative effect on human cancer cells. Uptake of SMoC-geminin was inhibited at 4 degrees C and by chlorpromazine, a compound that induces misassembly of clathrin-coated pits at the cell surface. Thus the mechanism of uptake is likely to be clathrin-mediated endocytosis.

Animals↗

PubMed2006

Fbxo7 gets proactive with cyclin D/cdk6.

Although all three D cyclins bind and activate cdks 2, 4 and 6, Fbxo7 has been characterised as a selective enhancer of cdk6 activity. It increases activation by directly facilitating cdk6 interaction with viral and cellular D cyclins. Fbxo7 overexpression has transforming activity in murine fibroblasts, and it is also highly expressed in human cancers, suggesting it is a potential oncogene. Fbxo7 has the ability to activate cell cycle regulators, and is part of an E3 ubiquitin ligase. We postulate Fbxo7 coordinates the ubiquitination of its substrates with cell cycle entry. It may therefore represent a means to integrate cell signals and control disparate biological processes during the early part of the cell cycle.

Animals↗

PubMed2005

Transforming activity of Fbxo7 is mediated specifically through regulation of cyclin D/cdk6.

D cyclins (D1, D2 and D3) and their catalytic subunits (cyclin-dependent kinases cdk4 and cdk6) have a facilitating, but nonessential, role in cell cycle entry. Tissue-specific functions for D-type cyclins and cdks have been reported; however, the biochemical properties of these kinases are indistinguishable. We report that an F box protein, Fbxo7, interacted with cellular and viral D cyclins and distinguished among the cdks that bind D-type cyclins, specifically binding cdk6, in vitro and in vivo. Fbxo7 specifically regulated D cyclin/cdk6 complexes: Fbxo7 knockdown decreased cdk6 association with cyclin and its overexpression increased D cyclin/cdk6 activity and E2F activity. Fbxo7 interacted with p27, but its enhancement of cyclin D/cdk6 activity was p21/p27 independent. Fbxo7 overexpression transformed murine fibroblasts, rendering them tumorigenic in athymic nude mice. Transformed phenotypes were dependent on cdk6, as knockdown of cdk6 reversed them. Fbxo7 was highly expressed in epithelial tumors, but not in normal tissues, suggesting that it may have a proto-oncogenic role in human cancers.

Active Transport, Cell Nucleus↗

PubMed2004

Regulation of growth signalling and cell cycle by Kaposi's sarcoma-associated herpesvirus genes.

Kaposi's sarcoma-associated herpesvirus (KSHV) is the primary aetiological agent of at least three malignancies associated with HIV infection and immunosuppression: Kaposi's sarcoma, primary effusion lymphoma and multicentric Castleman's disease. KSHV encodes proteins that deregulate key checkpoints in the signalling pathways governing cell proliferation, which may ultimately contribute to the virus' oncogenic potential. To alter cellular signalling associated with proliferation, these viral proteins function like growth factor ligands/receptors, signal transduction proteins, transcription factors and cell cycle regulators. This review focuses on the mechanisms by which some KSHV-encoded proteins activate signalling pathways and cell proliferation and their role in the pathogenesis of KSHV-driven mechanisms.

Cell Cycle↗

PubMed2003

RNA interference: a potential tool against Kaposi's sarcoma-associated herpesvirus.

PURPOSE OF REVIEW: RNA interference is a conserved cellular function that controls viral infection, the expression of transposable elements, repetitive sequences and genes in embryonic development. Originally described as an antiviral mechanism in plants, known as posttranscriptional gene silencing, it is now appreciated that this phenomenon occurs in all living cells. Double-stranded RNA, when acting as part of RNA interference, reduces expression of genes with sequence similarity, but has no effect on the expression of genes of unrelated sequence. Studies of RNA interference in mammalian cells have demonstrated that exogenous genes delivered by DNA transfection as well as endogenous gene expression can be suppressed by the delivery of RNA interference. We discuss here the potential for exploiting this phenomenon to prevent or treat viral infections, in particular Kaposi's sarcoma-associated herpesvirus. RECENT FINDINGS: There have been several studies showing that RNA interference can be exploited to target a wide range of human viruses, including HIV-1, human T cell leukaemia virus-1, human papillomavirus, hepatitis B, hepatitis C and the polio virus. RNA interference is effective in mammalian cells and can be delivered by various methods. Double-stranded RNA has been injected into the tail veins of mice to block both virally and chemically induced hepatitis. SUMMARY: A greater understanding of RNA interference allows us to exploit this phenomenon in order to study the functions of genes in mammalian cells, and also to target the expression of mutated cellular or viral genes. New delivery techniques should be developed to allow the process to be used as a therapeutic tool against viruses and malignancies in humans.

Antiretroviral Therapy, Highly Active↗

PubMed2002

Distinct roles for cyclins E and A during DNA replication complex assembly and activation.

Initiation of DNA replication is regulated by cyclin-dependent protein kinase 2 (Cdk2) in association with two different regulatory subunits, cyclin A and cyclin E (reviewed in ref. 1). But why two different cyclins are required and why their order of activation is tightly regulated are unknown. Using a cell-free system for initiation of DNA replication that is based on G1 nuclei, G1 cytosol and recombinant proteins, we find that cyclins E and A have specialized roles during the transition from G0 to S phase. Cyclin E stimulates replication complex assembly by cooperating with Cdc6, to make G1 nuclei competent to replicate in vitro. Cyclin A has two separable functions: it activates DNA synthesis by replication complexes that are already assembled, and it inhibits the assembly of new complexes. Thus, cyclin E opens a 'window of opportunity' for replication complex assembly that is closed by cyclin A. The dual functions of cyclin A ensure that the assembly phase (G1) ends before DNA synthesis (S) begins, thereby preventing re-initiation until the next cell cycle.

Cell Cycle Proteins↗