Epstein-Barr virus (EBV) infection of primary B cells causes B-

The Kaposi's sarcoma-associated herpesvirus (KSHV) latency-associated nuclear antigen (LANA) protein is functionally pleiotropic. LANA contributes to KSHV-associated pathogenesis, in part, by increasing entry of cells into S phase through a process that is driven by LANA interaction with the serine-threonine kinase glycogen synthase kinase 3 (GSK-3) and stabilization of ␤-catenin. Kaposi's sarcoma-associated herpesvirus (KSHV) was discovered in lesions of Kaposi's sarcoma using differential display (12) and was subsequently recognized to also be associated with primary effusion lymphoma and multicentric Castleman's disease (10, 18, 52, 59). The KSHV latency-associated nuclear antigen (LANA) is one of a limited number of KSHV genes consistently expressed in latently infected cells and in KSHV-associated malignancies (47). LANA is encoded by KSHV ORF73 and has unique N-terminal and C-terminal domains separated by three sets of repeated sequences that represent approximately half of the total protein sequence. These repeats function similarly to the central repeat region of the Epstein-Barr virus EBNA-1 protein by inhibiting antigen presentation and allowing tumor cells expressing LANA to escape immune surveillance (2,16,70).LANA is a multifunctional protein that is essential for the replication (5, 20, 29, 34) and maintenance (4) of KSHV episomal DNA during latent infection. LANA binds to the terminal repeats of the KSHV genome (14, 25); links the genomes to the cell chromosomes through interactions with chromatinassociated proteins such as the core histones H2A and H2B, DEK, HP1, Brd4, and MeCP2 (6,28,37,69); and recruits cellular DNA replication machinery to the terminal repeats (45,60,62,64). Expression of LANA in a transgenic mouse generated activated, hyperproliferative B cells, and mice developed lymphoma with a long latency (19). LANA has multiple properties that could contribute to tumorigenesis. These include inhibition of p53-mediated apoptosis (9, 21), stimulation of S-phase entry through stabilization of ␤-catenin and upregulation of cyclin D1 (24) and through induction of Rb/ E2F-regulated genes (1, 49), and overcoming G 1 cell cycle arrest mediated by p16 (1) and BRD4 and BRD2 (46). LANA is also responsible for promoting KSHV latency gene expression at the expense of lytic induction and for some of the reprogramming of cell gene expression that occurs in KSHVinfected cells (1,57,65,66). Targeting of LANA to DNA either through the use of Gal4-LANA fusion proteins (38, 53) through binding of LANA to the KSHV terminal repeats (25) or through LANA recruitment to cell (57) or viral promoters (39, 42) leads to transcriptional repression. LANA binds to histone deacetylase-associated corepressors (38) and is also capable of recruiting de novo DNA methyltransferases and the histone methyl transferase SUV39H1 to downregulate targeted cell promoters through CpG methylation (50, 57).LANA has also been reported to increase expression of genes regulated by a variety of transcription factors (40,44,61,63). A source...

The Kaposi's sarcoma-associated herpesvirus latency-associated nuclear antigen (LANA) protein interacts with glycogen synthase kinase 3 (GSK-3) and relocalizes GSK-3 in a manner that leads to stabilization of ␤-catenin and upregulation of ␤-catenin-responsive cell genes. The LANA-GSK-3 interaction was further examined to determine whether there were additional downstream consequences. In the present study, the nuclear GSK-3 bound to LANA in transfected cells and in BCBL1 primary effusion lymphoma cells was found to be enriched for the inactive serine 9-phosphorylated form of GSK-3. The mechanism of inactivation of nuclear GSK-3 involved LANA recruitment of the extracellular signal-regulated kinases 1 and 2 (ERK1/2) and the ribosomal S6 kinase 1 (RSK1). ERK1/2 and RSK1 coprecipitated with LANA, and LANA was a substrate for ERK1 in vitro. A model is proposed for the overall inactivation of nuclear GSK-3 that incorporates the previously described GSK-3 phosphorylation of LANA itself. Functional inactivation of nuclear GSK-3 was demonstrated by the ability of LANA to limit phosphorylation of the known GSK-3 substrates C/EBP␤ and C/EBP␣. The effect of LANA-mediated ablation of C/EBP phosphorylation on differentiation was modeled in the well-characterized 3T3L1 adipogenesis system. LANA-expressing 3T3L1 cells were impaired in their ability to undergo differentiation and adipogenesis. C/EBP␤ induction followed the same time course as that seen in vector-transduced cells, but there was delayed and reduced induction of C/EBP␤ transcriptional targets in LANA-expressing cells. We conclude that LANA inactivates nuclear GSK-3 and modifies the function of proteins that are GSK-3 substrates. In the case of C/EBPs, this translates into LANA-mediated inhibition of differentiation.The latency-associated nuclear antigen (LANA) protein is one of a very limited number of Kaposi's sarcoma-associated herpesvirus (KSHV) proteins synthesized during KSHV latency and in the KSHV-associated tumors of Kaposi's sarcoma, primary effusion lymphoma (PEL), and multicentric Castleman's disease (25,41,52). LANA is encoded by KSHV open reading frame 73 and is expressed in latently infected cells from alternatively spliced transcripts that also encode the FLICE inhibitory protein v-FLIP and the D-type cyclin v-cyclin (12, 51, 57). A second downstream promoter that is responsive to the KSHV lytic transactivator known as the replication and transcription activator (RTA) also drives LANA expression (35,56), and this promoter may be important for the synthesis of LANA in cells immediately following KSHV infection (27). LANA is a multifunctional protein with DNA replication, chromosome tethering, antiapoptotic, cell cycle regulatory, and gene regulatory functions. LANA binds to sequences in the KSHV terminal repeats (8,19,22,23) that are essential for replication of the episomal KSHV genomes during latency (13,21). LANA also supports long-term episomal maintenance through the tethering of KSHV genomes to chromosomes (2,4,43). Interactions between the LAN...

After a reduction in full-time equivalents, 2 libraries in large teaching hospitals and 2 libraries in small community hospitals in a western US statewide health system saw opportunity for expansion through a regional reorganization. Despite a loss of 2/3 of the professional staff and a budgetary decrease of 27% over the previous 3 years, the libraries were able to grow business, usage, awareness, and collections through organizational innovation and improved efficiency. This paper describes the experience--including process, challenges, and lessons learned--of an organizational shift to regionalized services, collections, and staffing. Insights from this process may help similar organizations going through restructuring.

When the four Providence Health & Services libraries in Oregon regionalized services and resources, the transition, which was originally met with apprehension from some library users, turned out to be a resounding success. Despite a loss of two-thirds of the professional staff and a decreased budget, the new regionalized library experienced an increase in business and recognition. While many factors contributed to the success, a creative marketing and outreach campaign was a key component. This column describes the steps taken to promote regionalized library reference services and online resources.