Growth cone MKK7 mRNA targeting regulates MAP1b-dependent microtubule bundling to control neurite elongation

Local mRNA translation in neurons has been mostly studied during axon guidance and synapse formation but not during initial neurite outgrowth. We performed a genome-wide screen for neurite-enriched mRNAs and identified an mRNA that encodes mitogen-activated protein kinase kinase 7 (MKK7), a MAP kinase kinase (MAPKK) for Jun kinase (JNK). We show that MKK7 mRNA localizes to the growth cone where it has the potential to be translated. MKK7 is then specifically phosphorylated in the neurite shaft, where it is part of a MAP kinase signaling module consisting of dual leucine zipper kinase (DLK), MKK7, and JNK1. This triggers Map1b phosphorylation to regulate microtubule bundling leading to neurite elongation. We propose a model in which MKK7 mRNA localization and translation in the growth cone allows for a mechanism to position JNK signaling in the neurite shaft and to specifically link it to regulation of microtubule bundling. At the same time, this uncouples activated JNK from its functions relevant to nuclear translocation and transcriptional activation.

Interference by the intracellular parasite Theileria parva with T-cell signal transduction pathways induces transformation and protection against apoptosis

The intracellular parasite Theileria parva transforms bovine T-lymphocytes, inducing uncontrolled proliferation. Upon infection, cells cease to require antigenic stimulation and exogenous growth factors to proliferate. Earlier studies have shown that pathways triggered via stimulation of the T-cell receptor are silent in transformed cells. This is reflected by a lack of phosphorylation of key signalling molecules and the fact that proliferation is not inhibited by immunosuppressants such as cyclosporin and ascomycin that target calcineurin. This suggests that the parasite bypasses the normal T-cells activation pathways to induce proliferation. Among the MAP-kinase pathways, ERK and p38 are silent, and only Jun N-terminal kinase is activated. This appears to suffice to induce constitutive activation of the transcription factor AP-1. More recently, it could be shown that the presence of the parasite in the host cell cytoplasm also induces constitutive activation of NF-kappaB, a transcription factor involved in proliferation and protection against apoptosis. Activation is effectuated by parasite-induced degradation of IkappaBs, the cytoplasmic inhibitors which sequester NF-kappaB in the cytoplasm. NF-kappaB activation is resistant to the antioxidant N-acetyl cysteine and a range of other reagents, suggesting that activation might occur in an unorthodox manner. Studies using inhibitors and dominant negative mutants demonstrate that the parasite activates a NF-kappaB-dependent anti-apoptotic mechanism that protects the transformed cell form spontaneous apoptosis and is essential for maintaining the transformed state of the parasitised cell.

Collagen I modulates growth and gene expression in prostate cancer cells

Prostate cancer is the second leading cause of male cancer-related deaths in the United States. Interestingly, prostate cancer preferentially metastasizes to skeletal tissue. Once in the bone microenvironment, advanced prostate cancer becomes highly resistant to therapeutic modalities. Several factors, such as extracellular matrix (ECM) components, have been implicated in the spread and propagation of prostatic carcinoma. In these studies, we have utilized the PC3 cell line, derived from a human bone metastasis, to investigate the influence of the predominant bone ECM protein, type I collagen, on prostate cancer cell proliferation and gene expression. We have also initiated the design and production of ribozymes to specific gene targets that may influence prostate cancer bone metastasis. ^ Our results demonstrate that PC3 cells rapidly adhere and spread on collagen I to a greater degree than on fibronectin (FN) or poly-L-lysine (PLL). Flow cytometry analysis reveals the presence of the α1, α2 and α3 collagen binding integrin subunits. The use of antibody function blocking studies reveals that PC3 cells can utilize α2β 1 and α3β1 integrins to adhere to collagen I. Once plated on collagen I, the cells exhibit increased rates of proliferation compared with cells plated on FN or tissue culture plastic. Additionally, cells plated on collagen I show increased expression of proteins associated with progression through G1 phase of the cell cycle. Inhibitor studies point to a role for phosphatidylinositol 3-kinase (PI3K), MAP kinase (MAPK), and p70 S6 kinase in collagen I-mediated PC3 cell proliferation and cyclin D1 expression. To further characterize the effect of type I collagen on prostate cancer bone metastasis, we utilized a cDNA microarray strategy to monitor type I collagen-mediated changes in gene expression. Results of this analysis revealed a gene expression profile reflecting the increased proliferation occurring on type I collagen. Microarray analysis also revealed differences in the expression of specific gene targets that may impact on prostate cancer metastasis to bone. ^ As a result of our studies on the interaction of prostate cancer cells and the skeletal ECM, we sought to develop novel molecular tools for future gene therapy of functional knockdown experiments. To this end, we developed a series of ribozymes directed against the α2 integrin and at osteopontin, a protein implicated in the metastasis of various cancers, including prostate. These ribozymes should facilitate the future study of the mechanism of prostate cancer cell proliferation, and disease progression occurring at sites of skeletal metastasis where a type I collagen-based environment predominates. ^ Together these studies demonstrate the involvement of bone ECM proteins on prostate cancer cell proliferation and suggest that they may play a significant role on the growth of prostate metastases once in the bone microenvironment. ^

Ser88 phosphorylation regulates dynein light chain 1 (DLC1) function in the mouse mammary gland

Dynein light chain 1 (DLC1) is a highly conserved and ubiquitously expressed protein which might have critical cellular function as total loss of DLC1 caused Drosophila embryonic death. Despite many proteins and RNAs interaction with it identified, DLC1's function(s) and regulation are largely unknown. Recently, DLC1 was identified as a physiological substrate of P21-activate kinase 1(Pak1) kinase from a human mammary cDNA library in a yeast-2-hybridization screening assay. Studies in primary human tumors and cell culture implicated that DLC1 could promote mammary cancerous phenotypes, and more importantly, Ser88 phosphorylation of DLC1by Pak1 kinase was found to be essential for DLC1's tumorigenic activities. Based on the above tissue culture studies, we hypothesized that Ser88 phosphorylation regulates DLC1. ^ To test this hypothesis, we generated two transgenic mouse models: MMTV-DLC1 and MMTV-DLC1-S88A mice with mammary specific expression of the DLC1 and DLC1-S88A cDNAs. Both of the transgenic mice mammary glands showed rare tumor incidence which indicated DLC1 alone may not be sufficient for tumorigenesis in vivo. However, these mice showed a significant alteration of mammary development. Mammary glands from the MMTV-DLC1 mice had hyperbranching and alveolar hyperplasia, with elevated cell proliferation. Intriguingly, these phenotypes were not seen in the mammary glands from the MMTV-S88A mice. Furthermore, while MMTV-DLC1 glands were normal during involution, MMTV-S88A mice showed accelerated mammary involution with increase apoptosis and altered expression of involution-associated genes. Further analysis of the MMTV-S88A glands showed they had increased steady state level of Bim protein which might be responsible for the early involution. Finally, our in vitro data showed that Ser88 phosphorylation abolished DLC1 dimer and consequently might disturb its interaction with Bim and destabilize Bim. ^ Collectively, our findings provided in vivo evidence that Ser88 phosphorylation of DLC1 can regulate DLC1's function. In addition, Ser88 phosphorylation might be critical for DLC1 dimer-monomer transition. ^

Estradiol and insulin cross -talk in breast cancer

Approximately 33% of clinical breast carcinomas require estrogens to proliferate. Epidemiological data show that insulin resistance and diabetes mellitus is 2–3 times more prevalent in women with breast cancer than those with benign breast lesions, suggesting a clinical link between insulin and estradiol. Insulin and estradiol have a synergistic effect on the growth of MCF7 breast cancer cells, and long-term estradiol treatment upregulates the expression of the key insulin signaling protein IRS-1. The goal of this study was to further define the mechanism(s) of cross-talk between insulin and estradiol in regulating the growth of breast cancer. Using MCF7 cells, acute treatment with insulin or estradiol alone was found to stimulate two activities associated with growth: Erk MAP kinase and PI 3-kinase. However, combined acute treatment had an antagonistic effect on both activities. Acute estradiol treatment inhibited the insulin-stimulated tyrosine phosphorylation of IRS-1 while increasing its serine phosphorylation; the serine phosphorylation was attenuated by the PI 3-kinase inhibitor wortmannin. The acute antagonism observed with combined estradiol and insulin are not consistent with the long-term synergistic effect on growth. In contrast, chronic estradiol treatment enhanced the insulin-sensitivity of breast cancer cells as measured by increases in total cellular insulin-stimulated tyrosine phosphorylation of IRS-1 and activation of PI 3-kinase. Estradiol stimulation of gene transcription was found to require PI 3-kinase activity but not MAP kinase activity. Insulin alone had no effect on ER transcriptional activity, but chronic treatment in combination with estradiol resulted in synergism of ER transcription. The synergistic effect of insulin and estradiol on MCF7 cell growth was also found to require PI 3-kinase but not MAP kinase activity. Therefore, chronic estradiol treatment increases insulin stimulation of PI 3-kinase, and PI 3-kinase is required for estradiol stimulation of gene transcription alone and in combined synergy with insulin. These data demonstrate that PI 3-kinase is the locus for the cross-talk between insulin and estradiol which results in enhanced breast cancer growth with long-term exposure to both hormones. This may have important clinical implications for women with high risk for breast cancer and/or diabetes mellitus. ^

Geological map of HANSSONHORNA, Heimefrontfjella, Antarctica (Scale 1:25,000)

Topographic data of this geological map were obtained through stereoscopic aerial photo interpretation. The photogrammetric photo flights were undertaken in 1986 by the Institut für Angewandte Geodäsie, Frankfurt. Horizontal ground control points required for aerial photo interpretation were determined by means of Doppler satellite observation during the 2nd German Neuschwabenland Expedition 1985/86. Vertical ground control points were taken from unpublished map drafts at 1:100 000 scale by Norsk Polarinstitutt, Oslo. The elevation above mean sea level was transferred to Heimefrontfjella barometrically. For this reason assertions concerning the absolute elevation (referred to sea level) are uncertain. Contours and spot heights presented on the map were obtained from the photogrammetric evaluation of the photography taken in 1986; relative elevation data (hight differences) are accurate to approximately ±10 m.

Geological map of RYGHNUTEN, Heimefrontfjella, Antarctica (Scale 1:25,000)

Topographic data of this geological map were obtained through stereoscopic aerial photo interpretation. The photogrammetric photo flights were undertaken in 1986 by the Institut für Angewandte Geodäsie, Frankfurt. Horizontal ground control points required for aerial photo interpretation were determined by means of Doppler satellite observation during the 2nd German Neuschwabenland Expedition 1985/86. Vertical ground control points were taken from unpublished map drafts at 1:100 000 scale by Norsk Polarinstitutt, Oslo. The elevation above mean sea level was transferred to Heimefrontfjella barometrically. For this reason assertions concerning the absolute elevation (referred to sea level) are uncertain. Contours and spot heights presented on the map were obtained from the photogrammetric evaluation of the photography taken in 1986; relative elevation data (hight differences) are accurate to approximately ±10 m.

Geological map of WORSFOLDFJELLET, Heimefrontfjella, Antarctica (Scale 1:25,000)

Topographic data of this geological map were obtained through stereoscopic aerial photo interpretation. The photogrammetric photo flights were undertaken in 1986 by the Institut für Angewandte Geodäsie, Frankfurt. Horizontal ground control points required for aerial photo interpretation were determined by means of Doppler satellite observation during the 2nd German Neuschwabenland Expedition 1985/86. Vertical ground control points were taken from unpublished map drafts at 1:100 000 scale by Norsk Polarinstitutt, Oslo. The elevation above mean sea level was transferred to Heimefrontfjella barometrically. For this reason assertions concerning the absolute elevation (referred to sea level) are uncertain. Contours and spot heights presented on the map were obtained from the photogrammetric evaluation of the photography taken in 1986; relative elevation data (height differences) are accurate to approximately ±10 m.

Geological map of JUCKESKAMMEN, Heimefrontfjella, Antarctica (Scale 1:25,000)

Topographic data of this geological map were obtained through stereoscopic aerial photo interpretation. The photogrammetric photo flights were undertaken in 1986 by the Institut für Angewandte Geodäsie, Frankfurt. Horizontal ground control points required for aerial photo interpretation were determined by means of Doppler satellite observation during the 2nd German Neuschwabenland Expedition 1985/86. Vertical ground control points were taken from unpublished map drafts at 1:100 000 scale by Norsk Polarinstitutt, Oslo. The elevation above mean sea level was transferred to Heimefrontfjella barometrically. For this reason assertions concerning the absolute elevation (referred to sea level) are uncertain. Contours and spot heights presented on the map were obtained from the photogrammetric evaluation of the photography taken in 1986; relative elevation data (hight differences) are accurate to approximately ±10 m.

Geological map of GRAMKROKEN, Heimefrontfjella, Antarctica (Scale 1:25,000)

Topographic data of this geological map were obtained through stereoscopic aerial photo interpretation. The photogrammetric photo flights were undertaken in 1986 by the Institut für Angewandte Geodäsie, Frankfurt. Horizontal ground control points required for aerial photo interpretation were determined by means of Doppler satellite observation during the 2nd German Neuschwabenland Expedition 1985/86. Vertical ground control points were taken from unpublished map drafts at 1:100 000 scale by Norsk Polarinstitutt, Oslo. The elevation above mean sea level was transferred to Heimefrontfjella barometrically. For this reason assertions concerning the absolute elevation (referred to sea level) are uncertain. Contours and spot heights presented on the map were obtained from the photogrammetric evaluation of the photography taken in 1986; relative elevation data (hight differences) are accurate to approximately ±10 m.

Geological map of KVITEBOTNEN, Heimefrontfjella, Antarctica (Scale 1:25,000)

Topographic data of this geological map were obtained through stereoscopic aerial photo interpretation. The photogrammetric photo flights were undertaken in 1986 by the Institut für Angewandte Geodäsie, Frankfurt. Horizontal ground control points required for aerial photo interpretation were determined by means of Doppler satellite observation during the 2nd German Neuschwabenland Expedition 1985/86. Vertical ground control points were taken from unpublished map drafts at 1:100 000 scale by Norsk Polarinstitutt, Oslo. The elevation above mean sea level was transferred to Heimefrontfjella barometrically. For this reason assertions concerning the absolute elevation (referred to sea level) are uncertain. Contours and spot heights presented on the map were obtained from the photogrammetric evaluation of the photography taken in 1986; relative elevation data (hight differences) are accurate to approximately ±10 m.

Geological map of SIRINUTEN, Heimefrontfjella, Antarctica (Scale 1:25,000)

Topographic data of this geological map were obtained through stereoscopic aerial photo interpretation. The photogrammetric photo flights were undertaken in 1986 by the Institut für Angewandte Geodäsie, Frankfurt. Horizontal ground control points required for aerial photo interpretation were determined by means of Doppler satellite observation during the 2nd German Neuschwabenland Expedition 1985/86. Vertical ground control points were taken from unpublished map drafts at 1:100 000 scale by Norsk Polarinstitutt, Oslo. The elevation above mean sea level was transferred to Heimefrontfjella barometrically. For this reason assertions concerning the absolute elevation (referred to sea level) are uncertain. Contours and spot heights presented on the map were obtained from the photogrammetric evaluation of the photography taken in 1986; relative elevation data (hight differences) are accurate to approximately ±10 m.

Geological map of NORUMNUTEN, Heimefrontfjella, Antarctica (Scale 1:25,000)

Topographic data of this geological map were obtained through stereoscopic aerial photo interpretation. The photogrammetric photo flights were undertaken in 1986 by the Institut für Angewandte Geodäsie, Frankfurt. Horizontal ground control points required for aerial photo interpretation were determined by means of Doppler satellite observation during the 2nd German Neuschwabenland Expedition 1985/86. Vertical ground control points were taken from unpublished map drafts at 1:100 000 scale by Norsk Polarinstitutt, Oslo. The elevation above mean sea level was transferred to Heimefrontfjella barometrically. For this reason assertions concerning the absolute elevation (referred to sea level) are uncertain. Contours and spot heights presented on the map were obtained from the photogrammetric evaluation of the photography taken in 1986; relative elevation data (hight differences) are accurate to approximately ±10 m.

Geological map of northern XU-Fjella, Heimefrontfjella, Dronning Maud Land, Antarctica (Scale 1:25,000)

The geological map shows the northeastern part of the polyphase deformed Sivorg Terrane in the Heimefrontfjella/Dronning Maud Land. The basement was affected by late Mesoproterozoic and Cambrian deformation and metamorphism. Geological mapping was carried out during the Antarctic Expedition 2000/01 of the Alfred Wegener Institute for Polar and Marine Research. Topographic data were obtained through stereoscopic aerial photo interpretation. The photogrammetric photo flights were undertaken in 1986 by the Institut für Angewandte Geodäsie, Frankfurt/M. Horizontal ground control points required for aerial photo interpretation were determined by means of Doppler satellite observation during the 2nd German Neuschwabenland Expedition 1985/86. Vertical ground control points were taken from unpublished map drafts at 1:100 000 scale by Norsk Polarinstitutt, Oslo. The elevation above mean sea level was transferred to Heimefrontfjella barometrically. For this reason assertions concerning the absolute elevation (referred to sea level) are uncertain. Contours and spot heights presented on the map were obtained from the photogrammetric evaluation of the photography taken in 1986; relative elevation data (height differences) are accurate to approximately ±10 m. Published by Fachbereich Geowissenschaften, Universität Bremen & Geologisches Institut, RWTH Aachen.

Geological map of HANSSONHORNA, Heimefrontfjella, Dronning Maud Land, Antarctica, 2nd revised Edition (Scale 1:25,000)

The geological map shows the border area between the polyphase (late Mesoproterozoic and Cambrian) deformed Sivorg Terrane and the Kottas Terrane where a pervasive Cambrian tectonometamorphic overprints is lacking. Geological revision mapping was carried out during the Antarctic Expedition 2000/01 of the Alfred Wegener Institute for Polar and Marine Research. Topographic data were obtained through stereoscopic aerial photo interpretation. The photogrammetric photo flights were undertaken in 1986 by the Institut für Angewandte Geodäsie, Frankfurt. Horizontal ground control points required for aerial photo interpretation were determined by means of Doppler satellite observation during the 2nd German Neuschwabenland Expedition 1985/86. Vertical ground control points were taken from unpublished map drafts at 1:100 000 scale by Norsk Polarinstitutt, Oslo. The elevation above mean sea level was transferred to Heimefrontfjella barometrically. For this reason assertions concerning the absolute elevation (referred to sea level) are uncertain. Contours and spot heights presented on the map were obtained from the photogrammetric evaluation of the photography taken in 1986; relative elevation data (height differences) are accurate to approximately ±10 m. Published by Geologisches Institut der RWTH Aachen & Fachbereich Geowissenschaften, Bremen.