Expression of LASS2 controlled by LAG1 or ADH1 promoters cannot functionally complement Lag1p.
LAG1 contributes to the substrate specificity and catalytic activity of ceramide synthases in Saccharomyces cerevisiae. Double deletion of LAG1 and its yeast homologue LAC1 results in the slow growth defect of the cell under certain genetic backgrounds. LASS2, containing the conserved TLC domain and the specific HOX domain, is a human homologue of Lag1p. In this study, shuffling tests and tetrad analyses were carried out to examine the complementation between Lag1p and LASS2 or its fragment containing the TLC domain but lacking the HOX domain (LASS2DeltaHOX). Controlled by either the natural weak LAG1 promoter or the strong yeast ADH1 promoter, LASS2 and LASS2DeltaHOX could not rescue the slow growth defect of double mutant. The results indicated that LASS2 or LASS2DeltaHOX could not functionally complement Lag1p.
Yu
Yao
Y
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, PR China.
Lu
Hong
H
Pan
Hui
H
Ma
Jun-Hao
JH
Ding
Zhi-Jie
ZJ
Li
Yu-Yang
YY
D000426:Alcohol Dehydrogenase / Q000235:genetics*; D000595:Amino Acid Sequence; D005816:Genetic Complementation Test; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*; D016415:Sequence Alignment; D051049:Sphingosine N-Acyltransferase; D013997:Time Factors; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*
C087436:LAG1 protein, S cerevisiae; D008565:Membrane Proteins; D029701:Saccharomyces cerevisiae Proteins; D025521:Tumor Suppressor Proteins; C509911:ADH2 protein, S cerevisiae; D000426:Alcohol Dehydrogenase; C438311:CERS2 protein, human; D051049:Sphingosine N-Acyltransferase