Human apolipoprotein A-IV: displacement from the surface of triglyceride-rich particles by HDL2-associated C-apoproteins

Human apolipoprotein A-IV rapidly dissociates from the surface of lymph chylomicrons following their entry into circulation by an unknown mechanism. We have therefore investigated the binding of human apoA-IV to triglyceride-rich particles and the interaction of these apoA-IV/lipid complexes with human HDL2. Human apoA-IV was purified from lipoprotein depleted serum (J. Lipid Res. 1983. 24:52-59). Triglyceride-rich particles of well-defined properties were isolated from Intralipid, a commercially available phospholipid-triglyceride emulsion. Various concentrations of radiolabeled human apoA-IV were incubated at 24 degrees C with a fixed quantity of lipid particles; the particles were reisolated by centrifugation, and bound and free apoA-IV were quantitated. In 50 mM Tris, pH 7.4, apoA-IV bound to the triglyceride-rich particles in a non-cooperative manner, with a Kd of 2.0 microM. The calculated maximal binding was 4.96 X 10(-4) mol of apoA-IV bound per mol of phospholipid. The addition of increasing amounts of human HDL2 to the incubations caused the progressive dissociation of apoA-IV from the triglyceride-rich particles. Analysis of the reisolated particles by isoelectric focusing demonstrated the presence of C-apoproteins, suggesting their transfer from HDL2. Addition of purified apoC-III-1 to the incubations at concentrations equivalent to those present in HDL2 caused a similar dissociation of apoA-IV. HDL2 was modified to selectively remove C-apoproteins, without alteration of other physical characteristics. This modified HDL2 was four times less effective in causing apoA-IV dissociation. These results demonstrate that the lipid binding properties of human apoA-IV may be quantitatively examined using triglyceride-rich particles as model chylomicrons. This approach reproduces in vitro the dissociation of apoA-IV that occurs in vivo when mesenteric lymph chylomicrons enter the circulation, and suggests that the primary mechanism for this phenomenon is the transfer of C-apoproteins from high density lipoproteins to the triglyceride-rich particle surface. We hypothesize that this mechanism may play an important role in the modulation of chylomicron apoA-IV content in man.     

DNA-fork displacement rates in cultured mammalian cells with mutations in ribonucleotide reductase

DNA-replication fork displacement rates were measured in mouse S49 lymphosarcoma cell lines and in derivatives of those cell lines. One of the derivatives lacks dCMP deaminase activity and two others bear defined mutations in ribonucleotide reductase. We also examined a revertant cell line that was selected from one of the ribonucleotide reductase mutants and has regained normal ribonucleotide reductase activity. Our results show a correlation between decreased fork-displacement rates and alterations in ribonucleotide reductase, suggesting a possible involvement of this enzyme in the replication apparatus.     

Expression cloning of the TGF-beta type II receptor, a functional transmembrane serine/threonine kinase.

A cDNA encoding the TGF-beta type II receptor protein has been isolated by an expression cloning strategy. The cloned cDNA, when transfected into COS cells, leads to overexpression of an approximately 80 kd protein that specifically binds radioiodinated TGF-beta 1. Excess TGF-beta 1 competes for binding of radioiodinated TGF-beta 1 in a dose-dependent manner and is more effective than TGF-beta 2. The predicted receptor structure includes a cysteine-rich extracellular domain, a single hydrophobic transmembrane domain, and a predicted cytoplasmic serine/threonine kinase domain. A chimeric protein containing the intracellular domain of the type II receptor and expressed in E. coli can phosphorylate itself on serine and threonine residues in vitro, indicating that the cytoplasmic domain of the type II receptor is a functional kinase. This result implicates serine/threonine phosphorylation as an important mechanism of TGF-beta receptor-mediated signaling.     

Activation of Ha-ras p21 by substitution, deletion, and insertion mutations.

The transforming activity of naturally arising ras oncogenes results from point mutations that affect residue 12 or 61 of the encoded 21-kilodalton protein (p21). By use of site-directed mutagenesis, we showed that deletions and insertions of amino acid residues in the region of residue 12 are also effective in conferring oncogenic activity on p21. Common to these various alterations is the disruption that they create in this domain of the protein, which we propose results in the inactivation of a normal function of the protein.     

<Emphasis Type="Italic">Paracoccidioides brasiliensis</Emphasis> Infection Mimicking Recurrent Hodgkin Lymphoma: A Case Report and Review of the Literature

Paracoccidioides infection is a rare entity in the USA. This dimorphic fungus is found in Central and South America and is thought to be acquired by inhalation through the soil. We report a case of Paracoccidioides brasiliensis infection presenting as a clavicular bone lesion, peripancreatic mass, and various skin lesions. A 35-year-old man with a history significant for Hodgkin lymphoma presented with a left clavicular mass that was suspected clinically and radiologically as recurrent Hodgkin lymphoma. He was not experiencing any associated symptoms and was undergoing chemotherapy treatment for his known Hodgkin disease. On CT imaging, the mass was seen as a lytic bone lesion with an overlying soft tissue mass. This was biopsied and histologically diagnosed as a Paracoccidioides brasiliensis infection with associated necrotizing granulomatous inflammation. Also found on the CT scan was an enlarging peripancreatic mass which on endoscopic biopsy had similar histologic findings. In conclusion, this report presents a rare case of Paracoccidioides brasiliensis infection mimicking recurrent Hodgkin lymphoma.     

Relationships among msx gene structure and function in zebrafish and other vertebrates

The zebrafish genome contains at least five msx homeobox genes, msxA, msxB, msxC, msxD, and the newly isolated msxE. Although these genes share structural features common to all Msx genes, phylogenetic analyses of protein sequences indicate that the msx genes from zebrafish are not orthologous to the Msx1 and Msx2 genes of mammals, birds, and amphibians. The zebrafish msxB and msxC are more closely related to each other and to the mouse Msx3. Similarly, although the combinatorial expression of the zebrafish msx genes in the embryonic dorsal neuroectoderm, visceral arches, fins, and sensory organs suggests functional similarities with the Msx genes of other vertebrates, differences in the expression patterns preclude precise assignment of orthological relationships. Distinct duplication events may have given rise to the msx genes of modern fish and other vertebrate lineages whereas many aspects of msx gene functions during embryonic development have been preserved.      

Characterization of tumors produced by signal peptide-basic fibroblast growth factor-transformed cells

Basic fibroblast growth factor (bFGF) is found in a variety of cells and tissues. We have previously shown that bFGF is a transforming growth factor, but only when fused to a signal peptide (sp-bFGF). Cells expressing the native bFGF are tumorigenic in nude mice only, where the tumors form at a low frequency and grow very slowly as compared to sp-bFGF tumors. The cells transformed by the sp-bFGF growth factor gene cause rapidly growing tumors within 10 days in 100% of syngeneic and nude mice. In nude mice, the tumors are highly vascularized, while the vascularization in immunocompetent syngeneic mice is not as prominent. The syngeneic mice have a characteristic humoral immune response to sp-bFGF tumors, which differs from that mounted against ras-induced tumors. The ability of bFGF to induce tumorigenicity is significant in view of the recent discoveries of three new oncogenes: hst, int-2, and an oncogene from a human colon cancer. In addition to homology with FGF, the proteins encoded by these oncogenes all have a potential signal peptide at the protein's amino terminus, suggesting a mode of action analogous to that of our artificial signal peptide-bFGF (sp-bFGF) transforming growth factor model system.