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41.
42.
Myosin heavy chain isoforms regulate muscle function but not myofibril assembly. 总被引:2,自引:1,他引:1 下载免费PDF全文
Myosin heavy chain (MHC) is the motor protein of muscle thick filaments. Most organisms produce many muscle MHC isoforms with temporally and spatially regulated expression patterns. This suggests that isoforms of MHC have different characteristics necessary for defining specific muscle properties. The single Drosophila muscle Mhc gene yields various isoforms as a result of alternative RNA splicing. To determine whether this multiplicity of MHC isoforms is critical to myofibril assembly and function, we introduced a gene encoding only an embryonic MHC into Drosophila melanogaster. The embryonic transgene acts in a dominant antimorphic manner to disrupt flight muscle function. The transgene was genetically crossed into an MHC null background. Unexpectedly, transformed flies expressing only the embryonic isoform are viable. Adult muscles containing embryonic MHC assemble normally, indicating that the isoform of MHC does not determine the dramatic ultrastructural variation among different muscle types. However, transformed flies are flightless and show reduced jumping and mating ability. Their indirect flight muscle myofibrils progressively deteriorate. Our data show that the proper MHC isoform is critical for specialized muscle function and myofibril stability. 相似文献
43.
Y. Pu I. A. Bernstein L. I. Bernstam R. L. Bronaugh 《In vitro cellular & developmental biology. Animal》1995,31(4):283-287
Summary The culture of cutaneous keratinocytes grown on a Puropore nylon microporous membrane at the air-liquid interface has been
shown to be similar to the epidermis in a number of molecular and morphologic characteristics but to exhibit a significantly
greater degree of tritiated water permeation. Various culture conditions have been altered in an effort to improve the water
barrier properties. A Kp value in the range of 5.5±1.6×10−3 has been obtained for 79% of the culturea) by plating 0.9×106 viable basal cells on a piece (13-mm diameter) of membrane for 7 days of submerged growth,b) by placing two membranes on two stacked glass fiber filters (47-mm extra-thick) in a culture dish (60 mm) for 14 days of
growth at the air-liquid interface,c) by replacing the growth medium, i.e., 1 ml of complete minimum essential medium (CMEM) every 24 h after lifting,d) by using 10% fetal bovine serum (FBS) in the CMEM during the submerged culture period and 15% FBS in the CMEM during the
lifted culture period, ande) by adding a dialysis membrane on top and a Puropore nylon membrane below the culture when the cultures were inserted in
the permeation cell for testing. The percentage of cultures with this value for Kp can be increased to 90% if only cultures
with yellow, smooth, and shiny surfaces are tested. This system should be useful as a replacement for skin in testing the
cutaneous permeation of some chemicals.
To whom correspondence should be addressed at 1528 Public Health, The University of Michigan, Ann Arbor, Michigan 48109-2029. 相似文献
44.
The Fps/Fes protein-tyrosine kinase promotes angiogenesis in transgenic mice. 总被引:11,自引:3,他引:8 下载免费PDF全文
P Greer J Haigh G Mbamalu W Khoo A Bernstein T Pawson 《Molecular and cellular biology》1994,14(10):6755-6763
45.
46.
Kinematics and Dynamics of Sorghum (Sorghum bicolor L.) Leaf Development at Various Na/Ca Salinities (I. Elongation Growth) 总被引:3,自引:1,他引:2 下载免费PDF全文
In many salt-sensitive species, elevated concentrations of Ca in the root growth media ameliorate part of the shoot growth reduction caused by NaCl stress. The physiological mechanisms by which Ca exerts protective effects on leaf growth are still not understood. Understanding growth inhibition caused by a stress necessitates locating the leaf expansion region and quantifying the profile of the growth reduction. This will enable comparisons and correlations with spatial gradients of probable physiologically inhibiting factors. In this work we applied the methods of growth kinematics to analyze the effects of elevated Ca concentrations on the spatial and temporal distributions of growth within the intercalary expanding region of salinized sorghum (Sorghum bicolor [L.] Moench, cv NK 265) leaves. NaCl (100 mM) caused a decrease in leaf elongation rate by shortening the leaf growing zone by 20%, as well as reducing the peak value of the longitudinal relative elemental growth rate (REG rate). Increasing the Ca concentrations from 1 to 10 mM restored the length of the growing zone of both emerged and unemerged salinized leaves and increased the peak value of the REG rate. The beneficial effects of supplemental Ca were, however, more pronounced in leaves after their appearance above the whorl of encircling older leaf sheaths. Elevated Ca then resulted in a peak value of REG rate higher than in the salinized leaves. The peak value of unemerged leaves was not increased, although it was maintained over a longer distance. The duration of elongation growth associated with a cell during its displacement from the leaf base was longer in salinized than control leaves, despite the fact that the elongation zone was shorter in salinity. Although partially restoring the length of the elongation region, supplemental Ca had no effect on the age of cessation of growth. Elongation of a tissue element, therefore, ceased when a cellular element reached a certain age and not a specific distance from the leaf base. 相似文献
47.
48.
H B Bernstein S P Tucker S R Kar S A McPherson D T McPherson J W Dubay J Lebowitz R W Compans E Hunter 《Journal of virology》1995,69(5):2745-2750
The transmembrane protein of human immunodeficiency virus type 1 (HIV-1) contains a leucine zipper-like (hydrophobic heptad) repeat which has been predicted to form an amphipathic alpha helix. To evaluate the potential of the hydrophobic heptad repeat to induce protein oligomerization, this region of gp41 has been cloned into the bacterial expression vector pRIT2T. The resulting plasmid, pRIT3, expresses a fusion protein consisting of the Fc binding domain of monomeric protein A, a bacterial protein, and amino acids 538 to 593 of HIV-1 gp41. Gel filtration chromatography demonstrated the presence of oligomeric forms of the fusion protein, and analytical centrifugation studies confirmed that the chimeric protein formed a higher-order multimer that was greater than a dimer. Thus, we have identified a region of HIV-1 gp41 which is capable of directing the oligomerization of a fusion protein containing monomeric protein A. Point mutations, previously shown to inhibit the biological activity of the HIV-1 envelope glycoprotein, have been engineered into the segment of gp41 contained in the fusion protein, and expressed mutant proteins were purified and analyzed via fast protein liquid chromatography. A point mutation in the heptad repeat, which changed the central isoleucine to an alanine, caused a significant (> 60%) decrease in oligomerization, whereas changing the central isoleucine to aspartate or proline resulted in almost a complete loss of oligomerization. Deletions of one, two, or three amino acids following the first isoleucine also resulted in a profound decrease in oligomerization. The inhibitory effects of the mutations on oligomer formation correlated with the effects of the same mutations on envelope glycoprotein-mediated fusion. A possible role of the leucine zipper-like region in the fusion process and in an oligomerization event distinct from assembly of the envelope glycoprotein complex is discussed. 相似文献
49.
50.
A spectrin-like protein from mouse brain membranes: phosphorylation of the 235,000-dalton subunit 总被引:2,自引:0,他引:2