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Dissecting the pet18mutation in Saccharomyces cervisiae: HTL1encodes a 7-kDa polypeptide that interacts with components of the RSC complex

Abstract

The yeast pet18mutant exhibits three distinct phenotypes: temperature-sensitive lethality, failure to maintain a dsRNA virus, and respiration deficiency. We have isolated a yeast mutant, H53, with phenotypes identical to those of pet18. Based on PCR and Southern hybridization analysis, H53 was found to result from a large chromosomal deletion extending from YCR019w to YCR028c on chromosome III. Genetic analysis was carried out on H53 to correlate individual loci with each of the observed phenotypes. Disruption of YCR020c-a/ MAK31 brought about a loss of dsRNA without affecting the temperature sensitive phenotype. The loss of YCR020w-b/ HTL1, which encodes a hypothetical protein of 78 amino acids in length, was shown to be responsible for the temperature-sensitive lethality of the H53 mutant. Using immunoblotting, we demonstrated that a 7-kDa protein was indeed expressed in wild-type yeast, but not in a HTL1deletion mutant. Moreover, the significance of HTL1was investigated by isolating genes that are functionally associated with HTL1. We demonstrated that Rsc8p interacts physically with Htl1p, and that the genes RSC3, STH1and RSC30interact with HTL1. Thus, HTL1may play a role in the function of the RSC complex.

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Acknowledgements

We thank Ms. Ue-Rong Chen for technical assistance, Dr. C. Holm for Ch1350 and pCH1122, Dr. J. J. Lin for the pRS426-based yeast genomic library and Dr. C. T. Chien for the yeast two-hybrid library. This work was supported by a grant from the National Science Council, Republic of China (NSC-87-2314-B-010-050)

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Correspondence to M. Y. Cheng.

Additional information

Communicated by C. P. Hollenberg

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Lu, Y., Lin, Y., Tsai, A. et al. Dissecting the pet18mutation in Saccharomyces cervisiae: HTL1encodes a 7-kDa polypeptide that interacts with components of the RSC complex. Mol Gen Genomics 269, 321–330 (2003). https://doi.org/10.1007/s00438-003-0835-1

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Keywords

  • HTL1
  • pet18
  • Temperature-sensitive lethality