Document Detail

Identification of a cDNA that encodes a 1-acyl-sn-glycerol-3-phosphate acyltransferase from Limnanthes douglasii.
MedLine Citation:
PMID:  7579178     Owner:  NLM     Status:  MEDLINE    
Two different techniques were used to isolate potential cDNAs for acyl-CoA: 1-acyl-sn-glycerol-3-phosphate acyltransferase (LPA-AT) enzymes from Limnanthes douglasii. Both heterologous screening with the maize pMAT1 clone and in vivo complementation of the Escherichia coli mutant JC201 which is deficient in LPA-AT activity, were carried out. Clones identified by these procedures were different. Homology searches demonstrated that the clone isolated by heterologous probing, pLAT1, encodes a protein which is most similar to the maize (open reading frame in pMAT1) and yeast SLC1 proteins, which are putative LPA-AT sequences. This L. douglasii sequence shows much lower homology to the E. coli LPA-AT protein PlsC, which is the only LPA-AT sequence confirmed by over-expression studies. The clone isolated by complementation, pLAT2, encodes a protein with homology to both SLC1 and PlsC. It was not possible to over-express the complementing protein encoded by pLAT2 but further experimentation on membranes from complemented JC201 demonstrated that they possess a substrate specificity distinctly different from PlsC and similar to Limnanthes sp. microsome specificity. This data strongly supports the contention that pLAT2 is an LPA-AT clone. Northern blot analysis revealed different expression patterns for the two genes in pLAT1 and pLAT2. Transcription of the gene encoding the insert of pLAT2 occurred almost exclusively in developing seed tissue, whilst the cDNA of pLAT1 hybridised to poly(A)+ mRNA from seed, stem and leaf, demonstrating more widespread expression throughout the plant. Southern blot analysis indicated that the cDNA of pLAT2 was transcribed from a single-copy gene while that for pLAT1 was a member of a small gene family.
A P Brown; C L Brough; J T Kroon; A R Slabas
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Plant molecular biology     Volume:  29     ISSN:  0167-4412     ISO Abbreviation:  Plant Mol. Biol.     Publication Date:  1995 Oct 
Date Detail:
Created Date:  1995-12-04     Completed Date:  1995-12-04     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9106343     Medline TA:  Plant Mol Biol     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  267-78     Citation Subset:  IM    
Department of Biological Sciences, University of Durham, UK.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/Z46836;  Z48730
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MeSH Terms
1-Acylglycerol-3-Phosphate O-Acyltransferase
Acyltransferases / genetics*,  metabolism
Amino Acid Sequence
Base Sequence
Blotting, Northern
Blotting, Southern
DNA, Complementary / genetics
Escherichia coli / genetics
Escherichia coli Proteins
Gene Library
Genetic Complementation Test
Microsomes / enzymology
Molecular Sequence Data
Open Reading Frames
Plants / enzymology,  genetics*
Sequence Analysis, DNA
Sequence Homology, Amino Acid
Substrate Specificity
Reg. No./Substance:
0/DNA, Complementary; 0/Escherichia coli Proteins; EC 2.3.-/Acyltransferases; EC O-Acyltransferase; EC protein, E coli

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