Regulation of coiled-coil assembly in tropomyosins

Esteban Araya, Christine Berthier, Edward Kim, Trevor Yeung, Xiaorong Wang, David M. Helfman

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Tropomyosins (TMs) are a family of actin filament-binding proteins. They consist of nearly 100% α-helix and assemble into parallel coiled-coil dimers. In vertebrates, TMs are encoded by four genes that give rise to at least 17 distinct isoforms through the use of alternative RNA splicing and alternative promoters. We have studied various aspects of the coiled-coil interactions among muscle and nonmuscle isoforms by the use of transfection of epitope-tagged constructs, followed by immunoprecipitation, SDS-PAGE, and Western blot analyses. For coiled-coil interactions between high-molecular-weight isoforms (284 amino acids), the information for homo- versus heterodimerization is contained in large part within the alternatively spliced exons of nonmuscle and muscle (skeletal and smooth) isoforms. Furthermore, sequences located in alternatively spliced exons encoding amino acids 39-80 (exons 2a/2b), amino acids 189-213 (exons 6a/6b), and amino acids 258-284 (exons 9a/9d) are critical for the selective formation of homo- versus heterodimers. Among low-molecular-weight isoforms (248 amino acids), TM-4 and TM-5 can form either homodimers or heterodimers. The trigger sequence (amino acids 190-202) is required for homodimerization of TM-4, but not heterodimerization of TM-4 with TM-5. How the dimeric state of TMs might play a role in their cellular localization and function is discussed.

Original languageEnglish (US)
Pages (from-to)176-183
Number of pages8
JournalJournal of Structural Biology
Volume137
Issue number1-2
DOIs
StatePublished - 2002
Externally publishedYes

Fingerprint

Tropomyosin
Exons
Protein Isoforms
Amino Acids
Molecular Weight
Microfilament Proteins
Alternative Splicing
Actin Cytoskeleton
Immunoprecipitation
Transfection
Smooth Muscle
Vertebrates
Epitopes
Polyacrylamide Gel Electrophoresis
Amino Acid Sequence
Skeletal Muscle
Western Blotting
Muscles

Keywords

  • Actin filaments
  • Coiled-coil
  • Cytoskeleton
  • Tropomyosin

ASJC Scopus subject areas

  • Structural Biology

Cite this

Araya, E., Berthier, C., Kim, E., Yeung, T., Wang, X., & Helfman, D. M. (2002). Regulation of coiled-coil assembly in tropomyosins. Journal of Structural Biology, 137(1-2), 176-183. https://doi.org/10.1006/jsbi.2002.4463

Regulation of coiled-coil assembly in tropomyosins. / Araya, Esteban; Berthier, Christine; Kim, Edward; Yeung, Trevor; Wang, Xiaorong; Helfman, David M.

In: Journal of Structural Biology, Vol. 137, No. 1-2, 2002, p. 176-183.

Research output: Contribution to journalArticle

Araya, E, Berthier, C, Kim, E, Yeung, T, Wang, X & Helfman, DM 2002, 'Regulation of coiled-coil assembly in tropomyosins', Journal of Structural Biology, vol. 137, no. 1-2, pp. 176-183. https://doi.org/10.1006/jsbi.2002.4463
Araya, Esteban ; Berthier, Christine ; Kim, Edward ; Yeung, Trevor ; Wang, Xiaorong ; Helfman, David M. / Regulation of coiled-coil assembly in tropomyosins. In: Journal of Structural Biology. 2002 ; Vol. 137, No. 1-2. pp. 176-183.
@article{73c5d05e80d2499d9dc7204822313ba8,
title = "Regulation of coiled-coil assembly in tropomyosins",
abstract = "Tropomyosins (TMs) are a family of actin filament-binding proteins. They consist of nearly 100{\%} α-helix and assemble into parallel coiled-coil dimers. In vertebrates, TMs are encoded by four genes that give rise to at least 17 distinct isoforms through the use of alternative RNA splicing and alternative promoters. We have studied various aspects of the coiled-coil interactions among muscle and nonmuscle isoforms by the use of transfection of epitope-tagged constructs, followed by immunoprecipitation, SDS-PAGE, and Western blot analyses. For coiled-coil interactions between high-molecular-weight isoforms (284 amino acids), the information for homo- versus heterodimerization is contained in large part within the alternatively spliced exons of nonmuscle and muscle (skeletal and smooth) isoforms. Furthermore, sequences located in alternatively spliced exons encoding amino acids 39-80 (exons 2a/2b), amino acids 189-213 (exons 6a/6b), and amino acids 258-284 (exons 9a/9d) are critical for the selective formation of homo- versus heterodimers. Among low-molecular-weight isoforms (248 amino acids), TM-4 and TM-5 can form either homodimers or heterodimers. The trigger sequence (amino acids 190-202) is required for homodimerization of TM-4, but not heterodimerization of TM-4 with TM-5. How the dimeric state of TMs might play a role in their cellular localization and function is discussed.",
keywords = "Actin filaments, Coiled-coil, Cytoskeleton, Tropomyosin",
author = "Esteban Araya and Christine Berthier and Edward Kim and Trevor Yeung and Xiaorong Wang and Helfman, {David M.}",
year = "2002",
doi = "10.1006/jsbi.2002.4463",
language = "English (US)",
volume = "137",
pages = "176--183",
journal = "Journal of Structural Biology",
issn = "1047-8477",
publisher = "Academic Press Inc.",
number = "1-2",

}

TY - JOUR

T1 - Regulation of coiled-coil assembly in tropomyosins

AU - Araya, Esteban

AU - Berthier, Christine

AU - Kim, Edward

AU - Yeung, Trevor

AU - Wang, Xiaorong

AU - Helfman, David M.

PY - 2002

Y1 - 2002

N2 - Tropomyosins (TMs) are a family of actin filament-binding proteins. They consist of nearly 100% α-helix and assemble into parallel coiled-coil dimers. In vertebrates, TMs are encoded by four genes that give rise to at least 17 distinct isoforms through the use of alternative RNA splicing and alternative promoters. We have studied various aspects of the coiled-coil interactions among muscle and nonmuscle isoforms by the use of transfection of epitope-tagged constructs, followed by immunoprecipitation, SDS-PAGE, and Western blot analyses. For coiled-coil interactions between high-molecular-weight isoforms (284 amino acids), the information for homo- versus heterodimerization is contained in large part within the alternatively spliced exons of nonmuscle and muscle (skeletal and smooth) isoforms. Furthermore, sequences located in alternatively spliced exons encoding amino acids 39-80 (exons 2a/2b), amino acids 189-213 (exons 6a/6b), and amino acids 258-284 (exons 9a/9d) are critical for the selective formation of homo- versus heterodimers. Among low-molecular-weight isoforms (248 amino acids), TM-4 and TM-5 can form either homodimers or heterodimers. The trigger sequence (amino acids 190-202) is required for homodimerization of TM-4, but not heterodimerization of TM-4 with TM-5. How the dimeric state of TMs might play a role in their cellular localization and function is discussed.

AB - Tropomyosins (TMs) are a family of actin filament-binding proteins. They consist of nearly 100% α-helix and assemble into parallel coiled-coil dimers. In vertebrates, TMs are encoded by four genes that give rise to at least 17 distinct isoforms through the use of alternative RNA splicing and alternative promoters. We have studied various aspects of the coiled-coil interactions among muscle and nonmuscle isoforms by the use of transfection of epitope-tagged constructs, followed by immunoprecipitation, SDS-PAGE, and Western blot analyses. For coiled-coil interactions between high-molecular-weight isoforms (284 amino acids), the information for homo- versus heterodimerization is contained in large part within the alternatively spliced exons of nonmuscle and muscle (skeletal and smooth) isoforms. Furthermore, sequences located in alternatively spliced exons encoding amino acids 39-80 (exons 2a/2b), amino acids 189-213 (exons 6a/6b), and amino acids 258-284 (exons 9a/9d) are critical for the selective formation of homo- versus heterodimers. Among low-molecular-weight isoforms (248 amino acids), TM-4 and TM-5 can form either homodimers or heterodimers. The trigger sequence (amino acids 190-202) is required for homodimerization of TM-4, but not heterodimerization of TM-4 with TM-5. How the dimeric state of TMs might play a role in their cellular localization and function is discussed.

KW - Actin filaments

KW - Coiled-coil

KW - Cytoskeleton

KW - Tropomyosin

UR - http://www.scopus.com/inward/record.url?scp=0036445587&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036445587&partnerID=8YFLogxK

U2 - 10.1006/jsbi.2002.4463

DO - 10.1006/jsbi.2002.4463

M3 - Article

C2 - 12064944

AN - SCOPUS:0036445587

VL - 137

SP - 176

EP - 183

JO - Journal of Structural Biology

JF - Journal of Structural Biology

SN - 1047-8477

IS - 1-2

ER -