ABSTRACT Microtubule-associated proteins can influence the organization, stability and dynamics of microtubules. We characterize a novel protein that associates with microtubules as assessed by immunofluorescence, immunoelectron microscopy, and co-sedimentation. The protein is expressed heavily in embryonic neurons and, to a lesser extent, in epithelial and mesodermal cells. The cDNA sequence predicts a protein of 1,547 amino acids and approximately 170 kDa. Immunoblot of embryo lysate demonstrates bands of approximately 240 and 260 kDa. The predicted amino acid sequence contains 77 potential serine/threonine phosphorylation sites. A distinctive feature is a predicted α-helical central domain comprising 21 identical repeats of an 11 amino acid sequence (PLEELRKDAAE). The protein is thermostable and has two major charge-domains: the amino-terminal 80% has an estimated pI of 4.0 and the carboxy-terminal 20%, a pI of 12.2. The protein shares several general biochemical and molecular features of MAPs, but its sequence is not similar to that of any described MAP.
A strong enhancer element is located within the long terminal repeats (LTRs) of exogenous, oncogenic avian retroviruses, such as Rous sarcoma virus (RSV) and the avian leukosis viruses. The LTRs of a second class of avian retroviruses, the endogenous viruses (evs), lack detectable enhancer function, a property that correlates with major sequence differences between the LTRs of these two virus groups. Despite this lack of independent enhancer activity, we previously identified sequences in ev LTRs that were able to functionally replace essential enhancer domains from the RSV enhancer with which they share limited sequence similarity. To identify candidate enhancer domains in ev LTRs that are functionally equivalent to those in RSV LTRs, we analyzed and compared ev and RSV LTR-specific DNA-protein interactions. Using this approach, we identified two candidate enhancer domains and one deficiency in ev LTRs. One of the proposed ev enhancer domains was identified as a CArG box, a motif also found upstream of several muscle-specific genes, and as the core sequence of the c-fos serum response element. The RSV LTR contains two CArG motifs, one at a previously identified site and one identified in this report at the same relative location as the ev CArG motif. A second factor binding site that interacts with a heat-stable protein was also identified in ev LTRs and, contrary to previous suggestions, appears to be different from previously described exogenous virus enhancer binding proteins. Finally, a deficiency in factor binding was found within the one inverted CCAAT box in ev LTRs, affirming the importance of sequences that flank CCAAT motifs in factor binding and providing a candidate defect in the ev enhancer.
Abstract: Great progress has been made over the last five years in our understanding of papillomavirus (PV) biology. New technology has enabled investigators to understand the relationship between the PV and its host. The PV cannot be cultured in vitro, and this has led to limitations for those wishing to study the biology of this virus. However, utilizing recombinant DNA technology, investigators now have abundant quantities of human papillomavirus (HPV) DNA for study. Such HPV genomes may be labeled with a radioisotope such as P 32 and used as a “probe” in hybridization studies to see if a given tissue contains HPV DNA. No longer are we limited to electron microscopy and immune studies in our efforts to identify HPV within benign or malignant tissues. Ultimately, we hope to understand the relationship between the virus and its host. This paper will concentrate on one aspect of this relationship—the immunology of HPV.
The cloning and partial characterization of the genome of human papillomavirus type 27 (HPV-27) is described. Hybridization analyses reveal that this is a new HPV type, with the strongest homology to HPV-2.
Human papillomavirus DNA has been detected in the semen of three patients, two of whom have severe chronic wart disease. These data support the contention that sexual transmission of human papillomavirus DNA could occur via semen, a possibility suggested by epidemiological data on the sexual transmission of human papillomavirus.
ABSTRACT Although the study of rostral-caudal segmentation of the insect body has been a rich source of information about embryonic pattern formation, relatively little is known of the process of proximal-distal segmentation of insect appendages. Here we demonstrate that during the period of limb segmentation, five segmentally iterated, sharply demarcated bands of cell surface alkaline phosphatase activity are expressed in embryonic grasshopper limbs. These bands span each intersegmental boundary in the limb as well as one boundary within the tarsus. Within appendages, expression is restricted to epithelial cells, where activity is present on both apical and basolateral surfaces. This epithelial alkaline phosphatase remains active at neutral pH, is insensitive to levamisole inhibition, and is strongly inhibited by nucleoside monophosphates. Treatment of embryos with phosphatidylinositol-specific phospholipase C releases almost all visible chromogenic activity, indicating that the epithelial alkaline phosphatase is anchored to the plasma membrane by glycosyl-phosphatidylinositol. When material released by phosphatidylinositol-specific phospholipase C is separated on native polyacrylamide gels, a single broad band of enzymatic activity is detected following incubation with substrate. A polyclonal antiserum raised against a 55×103Mr alkaline phosphatase from shrimp recognizes a single band of 56×103Mr on immunoblots of grasshopper membrane proteins. The spatially restricted expression of epithelial alkaline phosphatase suggests that it may be involved in epithelial cell rearrangements or shape changes associated with limb segmentation and morphogenesis. It also may contribute to definition of axon routes in the limb, since pioneer afferent growth cones turn at, and migrate along, the edge of one alkaline phosphatase-expressing epithelial domain.
