mRNA 3′ End Processing Factors: A Phylogenetic Comparison

Almost all eukaryotic mRNAs possess 3′ ends with a polyadenylate (poly(A)) tail. This poly(A) tail is not encoded in the genome but is added by the process of polyadenylation. Polyadenylation is a two-step process, and this process is accomplished by multisubunit protein factors. Here, we comprehensively compare the protein machinery responsible for polyadenylation of mRNAs across many evolutionary divergent species, and we have found these protein factors to be remarkably conserved in nature. These data suggest that polyadenylation of mRNAs is an ancient process

A large-scale analysis of mRNA polyadenylation of human and mouse genes

mRNA polyadenylation is a critical cellular process in eukaryotes. It involves 3′ end cleavage of nascent mRNAs and addition of the poly(A) tail, which plays important roles in many aspects of the cellular metabolism of mRNA. The process is controlled by various cis-acting elements surrounding the cleavage site, and their binding factors. In this study, we surveyed genome regions containing cleavage sites [herein called poly(A) sites], for 13 942 human and 11 155 mouse genes. We found that a great proportion of human and mouse genes have alternative polyadenylation (∼54 and 32%, respectively). The conservation of alternative polyadenylation type or polyadenylation configuration between human and mouse orthologs is statistically significant, indicating that alternative polyadenylation is widely employed by these two species to produce alternative gene transcripts. Genes belonging to several functional groups, indicated by their Gene Ontology annotations, are biased with respect to polyadenylation configuration. Many poly(A) sites harbor multiple cleavage sites (51.25% human and 46.97% mouse sites), leading to heterogeneous 3′ end formation for transcripts. This implies that the cleavage process of polyadenylation is largely imprecise. Different types of poly(A) sites, with regard to their relative locations in a gene, are found to have distinct nucleotide composition in surrounding genomic regions. This large-scale study provides important insights into the mechanism of polyadenylation in mammalian species and represents a genomic view of the regulation of gene expression by alternative polyadenylation

Alternative polyadenylation of cyclooxygenase-2

A biologically important human gene, cyclooxygenase-2 (COX-2), has been proposed to be regulated at many levels. While COX-1 is constitutively expressed in cells, COX-2 is inducible and is upregulated in response to many signals. Since increased transcriptional activity accounts for only part of the upregulation of COX-2, we chose to explore other RNA processing mechanisms in the regulation of this gene. We performed a comprehensive bioinformatics survey, the first of its kind known for human COX-2, which revealed that the human COX-2 gene has alternative polyadenylation (proximal and distal sites) and suggested that use of the alternative polyadenylation signals has tissue specificity. We experimentally established this in HepG2 and HT29 cells. We used an in vivo polyadenylation assay to examine the relative strength of the COX-2 proximal and distal polyadenylation signals, and have shown that the proximal polyadenylation signal is much weaker than the distal one. The efficiency of utilization of many suboptimal mammalian polyadenylation signals is affected by sequence elements located upstream of the AAUAAA, known as upstream efficiency elements (USEs). Here, we used in vivo polyadenylation assays in multiple cell lines to demonstrate that the COX-2 proximal polyadenylation signal contains USEs, mutation of the USEs substantially decreased usage of the proximal signal, and that USE spacing relative to the polyadenylation signal was significant. In addition, mutation of the COX-2 proximal polyadenylation signal to a more optimal sequence enhanced polyadenylation efficiency 3.5-fold. Our data suggest for the first time that alternative polyadenylation of COX-2 is an important post-transcriptional regulatory event

Transnational Legal Practice 2006-2007

Law practice continues to expand across borders, and lawyers and law firms from the United States and other countries are substantially invested in representations that take them outside of their home jurisdictions.[1] Unfortunately, reliable information relating to the extent of internationalization of the legal market is scarce. Neither the number of lawyers and law firms working in the international legal services market nor the receipts generated from internationally-related work are readily and reliably available. Nevertheless, statistics from both the United States and United Kingdom provide a sense of the numbers from the largest present sources of international legal practice. In the category of outbound services, for example, we can consider how U.S. lawyers and law firms serve foreign clients and U.S.-based clients in their offshore activities. One measure of these services could include the offshore activity of U.S. law firms. The American Lawyer Global 100 includes nine U.S.-based law firms with more than a quarter of their lawyers stationed outside of the United States, three of which support more than 50 percent of their lawyers working from overseas offices.[2] Another study of approximately sixty large U.S. law firms reported that those firms support approximately 375 offices overseas, where approximately 8,000 lawyers are working;[3] three-quarters of these lawyers are working in offices located in Europe. The U.S. Department of Commerce Bureau of Economic Analysis estimates that the export of legal services from the United States generated $4.3 billion in receipts in 2005, while imports of legal services were valued at$914 million, yielding a 4:1 surplus for balance-of-payment accounts.[4] According to the U.K. Department of Constitutional Affairs, British law firms generated £1.9 billion in exports in 2003, compared to £1.5 billion in imports.[5

Transnational Legal Practice 2006-07

This article reviews developments in transnational legal practice during 2006 and 2007, including international developments, U.S. developments and regional developments in Australia and Europe. The primary focus of the international developments section is the WTO\u27s General Agreement on Trade in Services (GATS). This article discusses GATS Track 1 Activities related to legal services, including the Legal Services Collective Requests and issues related to GATS Track 2 and the potential development of GATS disciplines. This section also surveys GATS-related initiatives of the American Bar Association and the International Bar Association and U.S. implementation of foreign lawyer multi-jurisdictional practice rules. In other areas, the international developments section addresses the development of a code of conduct for defense counsel practicing before the International Criminal Court and developments in the Financial Action Task Force (FATF). With respect to U.S. transnational legal practice developments, the article reviews U.S. bilateral free trade initiatives, lawyer discipline cooperation initiatives and significant litigation. The regional developments section documents the emergence in Australia of the first publicly-traded law firm and Australia\u27s efforts to promote greater multijurisdictional practice for Australian lawyers in the U.S. This section also reviews various European developments, including European competition law initiatives, the Akzo Nobel case currently pending before the European Court of Justice, and developments related to the free movement of lawyers, codes of conduct, money laundering and lawyer education

Transnational Legal Practice 2006-07

This article reviews developments in transnational legal practice during 2006 and 2007, including international developments, U.S. developments and regional developments in Australia and Europe. The primary focus of the international developments section is the WTO\u27s General Agreement on Trade in Services (GATS). This article discusses GATS Track 1 Activities related to legal services, including the Legal Services Collective Requests and issues related to GATS Track 2 and the potential development of GATS disciplines. This section also surveys GATS-related initiatives of the American Bar Association and the International Bar Association and U.S. implementation of foreign lawyer multi-jurisdictional practice rules. In other areas, the international developments section addresses the development of a code of conduct for defense counsel practicing before the International Criminal Court and developments in the Financial Action Task Force (FATF). With respect to U.S. transnational legal practice developments, the article reviews U.S. bilateral free trade initiatives, lawyer discipline cooperation initiatives and significant litigation. The regional developments section documents the emergence in Australia of the first publicly-traded law firm and Australia\u27s efforts to promote greater multijurisdictional practice for Australian lawyers in the U.S. This section also reviews various European developments, including European competition law initiatives, the Akzo Nobel case currently pending before the European Court of Justice, and developments related to the free movement of lawyers, codes of conduct, money laundering and lawyer education

miR-21-mediated regulation of 15-hydroxyprostaglandin dehydrogenase in colon cancer

This work is licensed under a Creative Commons Attribution 4.0 International License.Elevated prostaglandin E2 (PGE2) levels are observed in colorectal cancer (CRC) patients, and this increase is associated with poor prognosis. Increased synthesis of PGE2 in CRC has been shown to occur through COX-2-dependent mechanisms; however, loss of the PGE2-catabolizing enzyme, 15-hydroxyprostaglandin dehydrogenase (15-PGDH, HPGD), in colonic tumors contributes to increased prostaglandin levels and poor patient survival. While loss of 15-PGDH can occur through transcriptional mechanisms, we demonstrate that 15-PGDH can be additionally regulated by a miRNA-mediated mechanism. We show that 15-PGDH and miR-21 are inversely correlated in CRC patients, with increased miR-21 levels associating with low 15-PGDH expression. 15-PGDH can be directly regulated by miR-21 through distinct sites in its 3′ untranslated region (3′UTR), and miR-21 expression in CRC cells attenuates 15-PGDH and promotes increased PGE2 levels. Additionally, epithelial growth factor (EGF) signaling suppresses 15-PGDH expression while simultaneously enhancing miR-21 levels. miR-21 inhibition represses CRC cell proliferation, which is enhanced with EGF receptor (EGFR) inhibition. These findings present a novel regulatory mechanism of 15-PGDH by miR-21, and how dysregulated expression of miR-21 may contribute to loss of 15-PGDH expression and promote CRC progression via increased accumulation of PGE2.NIH R01 CA134609NIH R01 AR069044NIH/NCI Cancer Center Support Grant (P30 CA168524)New Jersey Commission on Cancer ResearchAmerican Heart Association (15GRNT23240019

Transnational Legal Practice 2008

This article reviews developments in transnational legal practice during 2006 and 2007, including international developments, U.S. developments and regional developments in Australia and Europe. The primary focus of the international developments section is the WTO\u27s General Agreement on Trade in Services (GATS). This article discusses GATS Track 1 Activities related to legal services, including the Legal Services Collective Requests and issues related to GATS Track 2 and the potential development of GATS disciplines. This section also surveys GATS-related initiatives of the American Bar Association and the International Bar Association and U.S. implementation of foreign lawyer multi-jurisdictional practice rules. In other areas, the international developments section addresses the development of a code of conduct for defense counsel practicing before the International Criminal Court and developments in the Financial Action Task Force (FATF). With respect to U.S. transnational legal practice developments, the article reviews U.S. bilateral free trade initiatives, lawyer discipline cooperation initiatives and significant litigation. The regional developments section documents the emergence in Australia of the first publicly-traded law firm and Australia\u27s efforts to promote greater multijurisdictional practice for Australian lawyers in the U.S. This section also reviews various European developments, including European competition law initiatives, the Akzo Nobel case currently pending before the European Court of Justice, and developments related to the free movement of lawyers, codes of conduct, money laundering and lawyer education

Transnational Legal Practice 2008

The current financial turmoil shaking the world illustrates the connectedness of national markets and economies. Legal practice is no exception: lawyers and their firms are experiencing the upheaval along with their clients.1 This has resulted in new opportunities for lawyers and firms–in bankruptcy and restructuring and, likely in the future, in regulatory advising as well–and, at the same time, in substantial challenges. The promise of benefits from a diversified practice–in terms of both substance and geography–is being tested as lawyers and law firms follow their clients through the uncertainties of the current economic conditions. As law firms cut the size of their legal and non-legal staffs and decrease compensation expectations, they also are capitalizing on the benefits of a geographically diverse footprint of practice by looking to overseas activities as opportunities for growth. The number of firms announcing new offices in the Middle East, for example, has not slowed during the economic crisis.2 Over the last twenty years or so, the growth of overseas activities of the largest U.S.-based law firms has far outpaced their growth within the United States, by a rate of ten-to-one.3 In 2007, more than 15,000 lawyers worked for the National Law Journal 250 firms in more than 550 offices located outside the United States.4 Indeed, two U.S.-based law firms with substantial investments in overseas offices joined the ranks of four of the London “Magic Circle” firms in a new category dubbed the “global elite.”5 But the description so far relates only to the most visible part of the story of the importance of overseas-related work for U.S. lawyers. Overseas-related work also supports lawyers working for firms that do not have formal international footprints. These may be firms with foreign clients or U.S.-based clients involved in offshore activities or partnerships. They may be firms that are members of international networks or associations of lawyers that serve as a source of referral relationships, among other things. Each of these arrangements points to the continuing importance of keeping watch over the regulatory and business environment for lawyers outside the United States. The U.S. Department of Commerce Bureau of Economic Analysis estimates that the export of U.S. legal services generated $6.4 billion in receipts in 2007, while imports of legal services were valued at nearly$1.6 billion, yielding a four-to-one surplus for balance-of-payment accounts.6 If globalization continues, as appears likely, lawyers may be able to rely on overseas activities as a sort of hedge against instability at home. Access to overseas legal markets, then, remains an issue of high priority

Transnational Legal Practice 2008

The current financial turmoil shaking the world illustrates the connectedness of national markets and economies. Legal practice is no exception: lawyers and their firms are experiencing the upheaval along with their clients.1 This has resulted in new opportunities for lawyers and firms–in bankruptcy and restructuring and, likely in the future, in regulatory advising as well–and, at the same time, in substantial challenges. The promise of benefits from a diversified practice–in terms of both substance and geography–is being tested as lawyers and law firms follow their clients through the uncertainties of the current economic conditions. As law firms cut the size of their legal and non-legal staffs and decrease compensation expectations, they also are capitalizing on the benefits of a geographically diverse footprint of practice by looking to overseas activities as opportunities for growth. The number of firms announcing new offices in the Middle East, for example, has not slowed during the economic crisis.2 Over the last twenty years or so, the growth of overseas activities of the largest U.S.-based law firms has far outpaced their growth within the United States, by a rate of ten-to-one.3 In 2007, more than 15,000 lawyers worked for the National Law Journal 250 firms in more than 550 offices located outside the United States.4 Indeed, two U.S.-based law firms with substantial investments in overseas offices joined the ranks of four of the London “Magic Circle” firms in a new category dubbed the “global elite.”5 But the description so far relates only to the most visible part of the story of the importance of overseas-related work for U.S. lawyers. Overseas-related work also supports lawyers working for firms that do not have formal international footprints. These may be firms with foreign clients or U.S.-based clients involved in offshore activities or partnerships. They may be firms that are members of international networks or associations of lawyers that serve as a source of referral relationships, among other things. Each of these arrangements points to the continuing importance of keeping watch over the regulatory and business environment for lawyers outside the United States. The U.S. Department of Commerce Bureau of Economic Analysis estimates that the export of U.S. legal services generated $6.4 billion in receipts in 2007, while imports of legal services were valued at nearly$1.6 billion, yielding a four-to-one surplus for balance-of-payment accounts.6 If globalization continues, as appears likely, lawyers may be able to rely on overseas activities as a sort of hedge against instability at home. Access to overseas legal markets, then, remains an issue of high priority