SAM domain-based protein oligomerization observed by live-cell fluorescence fluctuation spectroscopy.

PloS One
Brian D SlaughterR Li

Abstract

Sterile-alpha-motif (SAM) domains are common protein interaction motifs observed in organisms as diverse as yeast and human. They play a role in protein homo- and hetero-interactions in processes ranging from signal transduction to RNA binding. In addition, mutations in SAM domain and SAM-mediated oligomers have been linked to several diseases. To date, the observation of heterogeneous SAM-mediated oligomers in vivo has been elusive, which represents a common challenge in dissecting cellular biochemistry in live-cell systems. In this study, we report the oligomerization and binding stoichiometry of high-order, multi-component complexes of (SAM) domain proteins Ste11 and Ste50 in live yeast cells using fluorescence fluctuation methods. Fluorescence cross-correlation spectroscopy (FCCS) and 1-dimensional photon counting histogram (1dPCH) confirm the SAM-mediated interaction and oligomerization of Ste11 and Ste50. Two-dimensional PCH (2dPCH), with endogenously expressed proteins tagged with GFP or mCherry, uniquely indicates that Ste11 and Ste50 form a heterogeneous complex in the yeast cytosol comprised of a dimer of Ste11 and a monomer of Ste50. In addition, Ste50 also exists as a high order oligomer that does not interact with ...Continue Reading

References

Oct 17, 1998·Journal of Biotechnology·R RiglerA Schnetz
Feb 5, 1999·Science·C D ThanosJ U Bowie
Jul 2, 1999·Biophysical Journal·Y ChenE Gratton
Nov 26, 1999·Proceedings of the National Academy of Sciences of the United States of America·P KaskK Gall
Feb 13, 2001·Human Molecular Genetics·J A McGrathH van Bokhoven
Apr 10, 2002·Molecular and Cellular Biology·Rachel E LamsonPeter M Pryciak
Dec 5, 2002·Current Opinion in Structural Biology·Nancy L ThompsonNoah W Allen
Jan 25, 2003·Methods : a Companion to Methods in Enzymology·Kirsten Bacia, Petra Schwille
Feb 28, 2003·Methods : a Companion to Methods in Enzymology·Elke Haustein, Petra Schwille
May 27, 2003·Journal of Dental Research·P N KantaputraJ A McGrath
Oct 17, 2003·Nature·Won-Ki HuhErin K O'Shea
Dec 9, 2003·Trends in Biochemical Sciences·Chongwoo A Kim, James U Bowie
Dec 16, 2003·Proceedings of the National Academy of Sciences of the United States of America·Yan ChenJoachim D Müller
May 15, 2004·Molecular and Cellular Biology·Cristina E TognonPoul H B Sorensen
Aug 26, 2004·Journal of Molecular Biology·Jamie J KwanLogan W Donaldson
Sep 7, 2004·Clinical and Experimental Dermatology·M T SahinS Oztürkcan
Dec 15, 2004·Biophysical Journal·Yan ChenJoachim D Müller
Feb 4, 2005·Protein Science : a Publication of the Protein Society·Surajit BhattacharjyaFeng Ni
Jun 2, 2005·Science's STKE : Signal Transduction Knowledge Environment·Feng Qiao, James U Bowie
Nov 22, 2005·Nature Methods·Nathan C ShanerRoger Y Tsien
Dec 13, 2005·Journal of Molecular Biology·Jamie J KwanLogan W Donaldson
Jan 25, 2006·Nature Methods·Kirsten BaciaPetra Schwille
Jun 17, 2006·The EMBO Journal·Kazuo TatebayashiHaruo Saito
Sep 19, 2006·Biophysical Journal·Lindsey N HillesheimJoachim D Müller
Nov 13, 2007·Biophysical Journal·Michelle A DigmanEnrico Gratton
Dec 14, 2007·Proceedings of the National Academy of Sciences of the United States of America·Brian D SlaughterRong Li

❮ Previous
Next ❯

Citations

May 12, 2009·Eukaryotic Cell·Jens BoesgerMaria Mittag
Dec 17, 2010·Molecular Biology of the Cell·Brian D Slaughter, Rong Li
Feb 12, 2011·The Biochemical Journal·Wing ChanEd Manser
Sep 26, 2014·PloS One·Ramaswamy RakshambikaiNarayanaswamy Srinivasan
Jan 1, 2013·Genes·Jeong Eun ParkRenping Zhou
May 2, 2018·Proceedings of the National Academy of Sciences of the United States of America·Courtney A HollenderChris Dardick
Jul 15, 2018·Scientific Reports·Valentin DunsingSalvatore Chiantia
Jan 12, 2010·Developmental Cell·Brian D SlaughterRong Li

❮ Previous
Next ❯

Methods Mentioned

BETA
fluorescence
GTPase
PCR

Software Mentioned

Zeiss Confocor - 3

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.