Mutations in the yeast Nhp6 protein can differentially affect its in Vivo functions

Michael Kruppa, David J Kolodrubetz

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Nhp6A and Nhp6B from S. cerevisiae are required for viability at 38°C because they are involved in transcription of SNR6 by RNA polymerase III. Nhp6A also represses transcription of NHP6B by RNA polymerase II. Nhp6 belongs to the HMG1 family, defined by an 80 amino acid DNA binding domain, which includes six highly conserved residues. These amino acids were mutated in Nhp6A and their affects on Nhp6 function were assessed in vivo. Surprisingly, most of the changes allowed Nhp6A to function normally in supporting growth at 38°C. However, six mutants had differential effects on in vivo function. Finally, two of the mutant proteins that did not restore Nhp6A function in vivo were shown to bind and bend DNA in vitro as well as wild type. Together, these results suggest that Nhp6 interacts with another protein(s) to carry out some of its biological functions and that this interaction might differ at promoters transcribed by RNA polymerase II versus RNA polymerase III.

Original languageEnglish (US)
Pages (from-to)1292-1299
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume280
Issue number5
DOIs
StatePublished - 2001

Fingerprint

RNA Polymerase III
Fungal Proteins
RNA Polymerase II
HMGB1 Protein
Amino Acids
Mutation
DNA
Mutant Proteins
Saccharomyces cerevisiae
Transcription
Growth
Proteins
In Vitro Techniques

Keywords

  • HMG1
  • Nhp6A
  • Nhp6B
  • Saccharomyces cerevisiae
  • Structure-function analysis
  • Transcription

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Mutations in the yeast Nhp6 protein can differentially affect its in Vivo functions. / Kruppa, Michael; Kolodrubetz, David J.

In: Biochemical and Biophysical Research Communications, Vol. 280, No. 5, 2001, p. 1292-1299.

Research output: Contribution to journalArticle

@article{7841628b3da1429d9b1145127ebd2dcc,
title = "Mutations in the yeast Nhp6 protein can differentially affect its in Vivo functions",
abstract = "Nhp6A and Nhp6B from S. cerevisiae are required for viability at 38°C because they are involved in transcription of SNR6 by RNA polymerase III. Nhp6A also represses transcription of NHP6B by RNA polymerase II. Nhp6 belongs to the HMG1 family, defined by an 80 amino acid DNA binding domain, which includes six highly conserved residues. These amino acids were mutated in Nhp6A and their affects on Nhp6 function were assessed in vivo. Surprisingly, most of the changes allowed Nhp6A to function normally in supporting growth at 38°C. However, six mutants had differential effects on in vivo function. Finally, two of the mutant proteins that did not restore Nhp6A function in vivo were shown to bind and bend DNA in vitro as well as wild type. Together, these results suggest that Nhp6 interacts with another protein(s) to carry out some of its biological functions and that this interaction might differ at promoters transcribed by RNA polymerase II versus RNA polymerase III.",
keywords = "HMG1, Nhp6A, Nhp6B, Saccharomyces cerevisiae, Structure-function analysis, Transcription",
author = "Michael Kruppa and Kolodrubetz, {David J}",
year = "2001",
doi = "10.1006/bbrc.2001.4273",
language = "English (US)",
volume = "280",
pages = "1292--1299",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "5",

}

TY - JOUR

T1 - Mutations in the yeast Nhp6 protein can differentially affect its in Vivo functions

AU - Kruppa, Michael

AU - Kolodrubetz, David J

PY - 2001

Y1 - 2001

N2 - Nhp6A and Nhp6B from S. cerevisiae are required for viability at 38°C because they are involved in transcription of SNR6 by RNA polymerase III. Nhp6A also represses transcription of NHP6B by RNA polymerase II. Nhp6 belongs to the HMG1 family, defined by an 80 amino acid DNA binding domain, which includes six highly conserved residues. These amino acids were mutated in Nhp6A and their affects on Nhp6 function were assessed in vivo. Surprisingly, most of the changes allowed Nhp6A to function normally in supporting growth at 38°C. However, six mutants had differential effects on in vivo function. Finally, two of the mutant proteins that did not restore Nhp6A function in vivo were shown to bind and bend DNA in vitro as well as wild type. Together, these results suggest that Nhp6 interacts with another protein(s) to carry out some of its biological functions and that this interaction might differ at promoters transcribed by RNA polymerase II versus RNA polymerase III.

AB - Nhp6A and Nhp6B from S. cerevisiae are required for viability at 38°C because they are involved in transcription of SNR6 by RNA polymerase III. Nhp6A also represses transcription of NHP6B by RNA polymerase II. Nhp6 belongs to the HMG1 family, defined by an 80 amino acid DNA binding domain, which includes six highly conserved residues. These amino acids were mutated in Nhp6A and their affects on Nhp6 function were assessed in vivo. Surprisingly, most of the changes allowed Nhp6A to function normally in supporting growth at 38°C. However, six mutants had differential effects on in vivo function. Finally, two of the mutant proteins that did not restore Nhp6A function in vivo were shown to bind and bend DNA in vitro as well as wild type. Together, these results suggest that Nhp6 interacts with another protein(s) to carry out some of its biological functions and that this interaction might differ at promoters transcribed by RNA polymerase II versus RNA polymerase III.

KW - HMG1

KW - Nhp6A

KW - Nhp6B

KW - Saccharomyces cerevisiae

KW - Structure-function analysis

KW - Transcription

UR - http://www.scopus.com/inward/record.url?scp=0034810879&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034810879&partnerID=8YFLogxK

U2 - 10.1006/bbrc.2001.4273

DO - 10.1006/bbrc.2001.4273

M3 - Article

VL - 280

SP - 1292

EP - 1299

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 5

ER -