Detection and localization of a chloroplast-encoded HU-like protein that organizes chloroplast nucleoids

Plant Cell. 2002 Jul;14(7):1579-89. doi: 10.1105/tpc.002717.

Abstract

Chloroplast DNA (cpDNA) is packed into discrete structures called chloroplast nucleoids (cp-nucleoids). The structure of cpDNA is thought to be important for its maintenance and regulation. In bacteria and mitochondria, histone-like proteins (such as HU and Abf2, respectively) are abundant and play important roles in DNA organization. However, a primary structural protein has yet to be found in cp-nucleoids. Here, we identified an abundant DNA binding protein from isolated cp-nucleoids of the primitive red alga Cyanidioschyzon merolae. The purified protein had sequence homology with the bacterial histone-like protein HU, and it complemented HU-lacking Escherichia coli mutants. The protein, called HC (histone-like protein of chloroplast), was encoded by a single gene (CmhupA) in the C. merolae chloroplast genome. Using immunofluorescence and immunoelectron microscopy, we demonstrated that HC was distributed uniformly throughout the entire cp-nucleoid. The protein was expressed constitutively throughout the cell and the chloroplast division cycle, and it was able to condense DNA. These results indicate that HC, a bacteria-derived histone-like protein, primarily organizes cpDNA into the nucleoid.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algal Proteins / genetics
  • Algal Proteins / metabolism*
  • Amino Acid Sequence
  • Bacterial Proteins / genetics
  • Chloroplasts / metabolism*
  • DNA, Chloroplast / genetics*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Escherichia coli / genetics
  • Genetic Complementation Test
  • Immunoblotting
  • Molecular Sequence Data
  • Mutation
  • Rhodophyta / genetics*
  • Rhodophyta / metabolism
  • Saccharomyces cerevisiae Proteins*
  • Sequence Homology, Amino Acid
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • ABF2 protein, S cerevisiae
  • Algal Proteins
  • Bacterial Proteins
  • DNA, Chloroplast
  • DNA-Binding Proteins
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • histone-like protein HU, bacteria

Associated data

  • GENBANK/AB071964