Faculty Collaboration Database

Structural elements required for the localization of ASH1 mRNA and of a green fluorescent protein reporter particle in vivo. Curr Biol 1999 Mar 25;9(6):333-6

Date

04/21/1999

Pubmed ID

10209102

DOI

10.1016/s0960-9822(99)80144-4

Scopus ID

2-s2.0-0033602387 (requires institutional sign-in at Scopus site)   168 Citations

Abstract

The sorting of the Ash1 protein to the daughter nucleus of Saccharomyces cerevisiae in late anaphase of the budding cycle correlates with the localization of ASH1 mRNA at the bud tip [1] [2]. Although the 3' untranslated region (3' UTR) of ASH1 is sufficient to localize a reporter mRNA, it is not necessary, a result which indicates that other sequences are involved [1]. We report the identification of three additional cis-acting elements in the coding region. Each element alone, when fused to a lacZ reporter gene, was sufficient for the localization of the lacZ mRNA reporter to the bud. A fine-structure analysis of the 3' UTR element showed that its function in mRNA localization did not depend on a specific sequence but on the secondary and tertiary structure of a minimal 118 nucleotide stem-loop. Mutations in the stem-loop that affect the localization of the lacZ mRNA reporter also affected the formation of the localization particles, in living cells, composed of a green fluorescent protein (GFP) complexed with lacZ-ASH1-3' UTR mRNA [3]. A specific stem-loop in the 3' UTR of the ASH1 mRNA is therefore required for both localization and particle formation, suggesting that complex formation is part of the localization mechanism. An analysis on one of the coding-region elements revealed a comparable stem-loop structure with similar functional requirements.

Author List

Chartrand P, Meng XH, Singer RH, Long RM

Author

Roy M. Long PhD Assistant Dean, Associate Professor in the Medical School Regional Campuses department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

3' Untranslated Regions
Anaphase
Base Sequence
Cell Nucleus
Cell Polarity
DNA-Binding Proteins
Fungal Proteins
Genes, Reporter
Molecular Sequence Data
Nucleic Acid Conformation
Plasmids
RNA, Fungal
RNA, Messenger
Recombinant Fusion Proteins
Repressor Proteins
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins
Species Specificity
Structure-Activity Relationship
Suppression, Genetic
Transcription Factors
Transformation, Genetic