A humanized yeast system to analyze cleavage of prelamin A by ZMPSTE24

Eric Spear, Rebecca F. Alford, Tim D. Babatz, Kaitlin M. Wood, Otto W. Mossberg, Kamsi Odinammadu, Khurts Shilagardi, Jeffrey J Gray, Susan Doris Michaelis

Research output: Contribution to journalArticle

Abstract

The nuclear lamins A, B, and C are intermediate filament proteins that form a nuclear scaffold adjacent to the inner nuclear membrane in higher eukaryotes, providing structural support for the nucleus. In the past two decades it has become evident that the final step in the biogenesis of the mature lamin A from its precursor prelamin A by the zinc metalloprotease ZMPSTE24 plays a critical role in human health. Defects in prelamin A processing by ZMPSTE24 result in premature aging disorders including Hutchinson Gilford Progeria Syndrome (HGPS) and related progeroid diseases. Additional evidence suggests that defects in prelamin A processing, due to diminished ZMPSTE24 expression or activity, may also drive normal physiological aging. Because of the important connection between prelamin A processing and human aging, there is increasing interest in how ZMPSTE24 specifically recognizes and cleaves its substrate prelamin A, encoded by LMNA. Here, we describe two humanized yeast systems we have recently developed to examine ZMPSTE24 processing of prelamin A. These systems differ from one another slightly. Version 1.0 is optimized to analyze ZMPSTE24 mutations, including disease alleles that may affect the function or stability of the protease. Using this system, we previously showed that some ZMPSTE24 disease alleles that affect stability can be rescued by the proteasome inhibitor bortezomib, which may have therapeutic implications. Version 2.0 is designed to analyze LMNA mutations at or near the ZMPSTE24 processing site to assess whether they permit or impede prelamin A processing. Together these systems offer powerful methodology to study ZMPSTE24 disease alleles and to dissect the specific residues and features of the lamin A tail that are required for recognition and cleavage by the ZMPSTE24 protease.

Original languageEnglish (US)
JournalMethods
DOIs
StateAccepted/In press - Jan 1 2019

Fingerprint

Yeast
Yeasts
Processing
Lamin Type A
Aging of materials
Alleles
Peptide Hydrolases
Lamin Type B
Progeria
Premature Aging
Intermediate Filament Proteins
Nuclear Matrix
Defects
Mutation
Forms (concrete)
Proteasome Inhibitors
prelamin A
Nuclear Envelope
Metalloproteases
Eukaryota

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Spear, E., Alford, R. F., Babatz, T. D., Wood, K. M., Mossberg, O. W., Odinammadu, K., ... Michaelis, S. D. (Accepted/In press). A humanized yeast system to analyze cleavage of prelamin A by ZMPSTE24. Methods. https://doi.org/10.1016/j.ymeth.2019.01.001

A humanized yeast system to analyze cleavage of prelamin A by ZMPSTE24. / Spear, Eric; Alford, Rebecca F.; Babatz, Tim D.; Wood, Kaitlin M.; Mossberg, Otto W.; Odinammadu, Kamsi; Shilagardi, Khurts; Gray, Jeffrey J; Michaelis, Susan Doris.

In: Methods, 01.01.2019.

Research output: Contribution to journalArticle

Spear, Eric ; Alford, Rebecca F. ; Babatz, Tim D. ; Wood, Kaitlin M. ; Mossberg, Otto W. ; Odinammadu, Kamsi ; Shilagardi, Khurts ; Gray, Jeffrey J ; Michaelis, Susan Doris. / A humanized yeast system to analyze cleavage of prelamin A by ZMPSTE24. In: Methods. 2019.
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