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Membrane morphogenesis from cloned fragments of bacteriophage PM2 DNA that contain the sp6.6 gene
Author(s) -
Armour Gary A.,
Brewer Gregory J.
Publication year - 1990
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.4.5.2407591
Subject(s) - morphogenesis , bacteriophage , dna , gene , biology , microbiology and biotechnology , genetics , escherichia coli
The formation of new membrane vesicles normally occurs during eukaryotic organellogenesis and maturation of bacteriophage PM2. This virus was studied as a simple model for membrane morphogenesis. Previous biochemical and genetic studies suggest that a major structural protein of PM2, sp6.6, is an integral membrane protein involved in viral membrane morphogenesis. To establish the necessity of sp6.6 in membrane formation, restriction fragments of PM2 that contained the sp6.6 coding sequence were cloned into several plasmid vectors for expression in Escherichia coli. A construction in pBR322 containing two Hin dIII fragments of PM2 DNA caused production of intracellular membrane vesicles of the same size as those produced in the course of natural infection of Alteromonas espejiana. Similar results were obtained with a smaller construct of Hin dII fragments in the plasmid vector pPL‐λ. Expression of sp6.6 was detected via incorporation of 35 S‐labeled methionine after SDS‐polyacrylamide gel electrophoresis and with a specific rabbit antiserum on immunoblots. Other constructs did not produce recognizable vesicles or sp6.6. These results are the first to suggest that a hydrophobic membrane protein can cause development of new membrane structure.— A rmour , G. A.; B rewer , G. J. Membrane morphogenesis from cloned fragments of bacteriophage PM2 DNA that contain the sp6.6 gene. FASEB J. 4: 1488‐1493; 1990.