Expression of alternatively spliced human latent transforming growth factor beta binding protein-1.

Latent transforming growth factor beta binding protein-1 (LTBP1) is important in regulating the localisation and activation of transforming growth factor beta(TGFbeta). Three forms of LTBP1 mRNA have previously been described, LTBP1L, LTBP1S and LTBPdelta53. Here, we have analysed the LTBP1 coding sequence and identified two other spliced forms, LTBP1delta55 and LTBP1delta41. LTBP1delta55 is a short form of LTBPIL which lacks 55 amino acids including two consensus N-glycosylation sites and LTBP1delta41 is a form of LTBP1 which lacks the 12th EGF-like repeat. Furthermore, sequencing of genomic clones showed that splicing to generate LTBP1L occurs using an intra-exonic 3' splice acceptor site in the first coding exon of LTBP1S and that LTBP1delta55 arises from the alternative use of an exonic 3' splice acceptor site at the end of the following intron. LTBP1delta41 arises from skipping the exon which encodes the 12th EGF-like repeat. LTBP1delta55 and LTBP1delta41 mRNA are expressed in a wide variety of human tissues but the proportions of each splice form vary in the tissues.

Loss of a consensus heparin binding site by alternative splicing of latent transforming growth factor-beta binding protein-1.

Latent transforming growth factor-beta binding protein-1 (LTBP-1), plays an important role in controlling localisation and activation of transforming growth factor-beta (TGF-beta). We show that alternative splicing generates a form of mRNA which lacks bases 1277-1435 (termed LTBP-1delta53). The 53 amino acids encoded by these bases include the eighth cysteine of the first cysteine repeat and a consensus heparin binding sequence. Sequencing of genomic clones showed that alternative splicing resulted from the use of an intra-exonic 3' splice acceptor site. The loss of the heparin binding site implies that LTBP-1delta53 will bind to the extracellular matrix less efficiently than LTBP-1.

The latent transforming growth factor beta binding protein (LTBP) family.

The transforming growth factor beta (TGFbeta) cytokines are a multi-functional family that exert a wide variety of effects on both normal and transformed mammalian cells. The secretion and activation of TGFbetas is regulated by their association with latency-associated proteins and latent TGFbeta binding proteins (LTBPs). Over the past few years, three members of the LTBP family have been identified, in addition to the protoype LTBP1 first sequenced in 1990. Three of the LTBP family are expressed in a variety of isoforms as a consequence of alternative splicing. This review summarizes the differences between the isoforms in terms of the effects on domain structure and hence possible function. The close identity between LTBPs and members of the fibrillin family, mutations in which have been linked directly to Marfan's syndrome, suggests that anomalous expression of LTBPs may be associated with disease. Recent data indicating that differential expression of LTBP1 isoforms occurs during the development of coronary heart disease is considered, together with evidence that modulation of LTBP function, and hence of TGFbeta activity, is associated with a variety of cancers.

Iron supplementation moderates but does not cure the Belgrade anemia.

Belgrade rats inherit microcytic, hypochromic anemia as an autosomal recessive trait (gene symbol b). Erythrocytes and tissue are iron deficient in the face of elevated TIBC (total iron binding capacity) and percent iron saturation; iron injections increased the number of erythrocytes but their appearance remained abnormal. We have investigated iron supplements to improve husbandry of b/b rats and to learn more about the underlying defect and its tissue distribution. Weekly i.m. (intramuscular) injections of iron-dextran (Imferon at 30 mg kg-1) improved the anemia but did not alter the red cell morphology. Certain diets also improved the health of b/b rats when compared to standard rat chows by the criteria of weight, survival to adulthood, hematology and reproduction. The critical nutritional factor turned out to be iron bioavailability, with ferrous iron added to the diet improving the health of Belgrade rats without affecting the underlying erythroid defect. Tissue iron measurements after dietary or parenteral supplementation confirmed the iron deficient status of untreated b/b rats and established that dietary ferrous iron partially relieved this deficiency, with injections leading to greater amounts of tissue iron. Serum iron and TIBC were also found to be elevated in untreated b/b rats, with dietary supplementation decreasing but not eliminating the elevation in TIBC. These studies indicate that iron supplements can improve the health of b/b rats without altering the underlying defect and also suggest that the mutation could alter iron uptake in the GI (gastrointestinal) tract.