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Yarrowia lipolytica SRP54 Homolog and Translocation of Kar2p
Author(s) -
LEE IN HYUNG,
OGRYDZIAK DAVID M.
Publication year - 1997
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/(sici)1097-0061(199705)13:6<499::aid-yea100>3.0.co;2-m
Subject(s) - biology , yarrowia , signal recognition particle , chromosomal translocation , rna , open reading frame , saccharomyces cerevisiae , biochemistry , gene , peptide sequence , ribosome
To investigate the role of Srp54p in protein translocation, the Yarrowia lipolytica SRP54 homolog was cloned. Sequencing revealed an open reading frame of 536 amino acids coding for a 57·2 kilodalton polypeptide with 55 to 57% sequence identity to Srp54ps of Saccharomyces cerevisiae , Schizosaccharomyces pombe , and mouse. Like these Srp54ps, Y. lipolytica Srp54p has an N‐terminal domain with a highly conserved GTP‐binding site and a methionine‐rich C‐terminal domain. Differing results regarding the essentiality of SRP subunits were obtained. SRP54 is important but not essential for growth, but it was reconfirmed that at least one SRP RNA gene is essential. Cells with SRP54 deleted grow about six times more slowly than wild type; faster‐growing colonies, still growing much slower than wild type, appeared quite frequently. In srp54 Δ cells, no untranslocated alkaline extracellular protease precursor was detected. Therefore, to develop another reporter molecule the Y. lipolytica KAR2 homolog was cloned and Kar2p antibodies were produced. For Kar2p an untranslocated precursor was detected in srp54 Δ but not in wild‐type cells, suggesting that its translocation was defective in the srp54Δ cells. These results confirm an in vivo role for SRP in protein translocation in Y. lipolytica , suggest that SRP RNA or an SRP core‐particle has functions not shared by Srp54p, and show that, as in S. cerevisiae and Sz. pombe , reporter molecules differ in their dependency on SRP for translocation. The SRP54 and KAR2 sequences have been deposited in GenBank under Accession Numbers U42418 and U63136.