Cloning and expression of Drosophila HP1 homologs from a mealybug, Planococcus citri

H Epstein, T C James, P B Singh

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

The mealybug chromosome cycle is one of the most dramatic examples of genomic imprinting known. In embryos that are to become male the entire paternal chromosome set becomes heterochromatic and inactive at the blastoderm stage, while the maternal set remains active and euchromatic. HP1 is a protein from Drosophila melanogaster, which binds preferentially to heterochromatin on polytene chromosomes and is likely to be a modifier of position effect variegation. This paper describes the isolation and sequencing of two cDNA clones encoding HP1 homologs from the mealybug, Planococcus citri. The protein product of the cDNA clone that was closer to HP1 in sequence was expressed as a fusion protein in Escherichia coli, and polyclonal rat antibodies were raised against it. Immunohistochemistry to mealybug squash preparations showed that this protein was a male-specific nuclear protein, but that it was not specifically associated with the heterochromatic set of chromosomes.

Original languageEnglish
Pages (from-to)463-74
Number of pages12
JournalJournal of Cell Science
Volume101 ( Pt 2)
Publication statusPublished - Feb 1992

Fingerprint

Drosophila
Organism Cloning
Chromosomes
Chromosomal Position Effects
Complementary DNA
Clone Cells
Blastoderm
Polytene Chromosomes
Genomic Imprinting
Drosophila Proteins
Cucurbita
Heterochromatin
Escherichia coli Proteins
Nuclear Proteins
Proteins
Embryonic Structures
Immunohistochemistry
Mothers
Antibodies
Drosophila arc protein

Keywords

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Chromosomal Proteins, Non-Histone
  • Cloning, Molecular
  • Drosophila melanogaster
  • Genetic Engineering
  • Hemiptera
  • Insect Hormones
  • Insect Proteins
  • Molecular Sequence Data
  • Multigene Family
  • Nuclear Proteins
  • Recombinant Fusion Proteins
  • Sequence Homology, Nucleic Acid
  • Sex Characteristics
  • Tissue Distribution
  • Journal Article
  • Research Support, Non-U.S. Gov't

Cite this

Cloning and expression of Drosophila HP1 homologs from a mealybug, Planococcus citri. / Epstein, H; James, T C; Singh, P B.

In: Journal of Cell Science, Vol. 101 ( Pt 2), 02.1992, p. 463-74.

Research output: Contribution to journalArticle

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abstract = "The mealybug chromosome cycle is one of the most dramatic examples of genomic imprinting known. In embryos that are to become male the entire paternal chromosome set becomes heterochromatic and inactive at the blastoderm stage, while the maternal set remains active and euchromatic. HP1 is a protein from Drosophila melanogaster, which binds preferentially to heterochromatin on polytene chromosomes and is likely to be a modifier of position effect variegation. This paper describes the isolation and sequencing of two cDNA clones encoding HP1 homologs from the mealybug, Planococcus citri. The protein product of the cDNA clone that was closer to HP1 in sequence was expressed as a fusion protein in Escherichia coli, and polyclonal rat antibodies were raised against it. Immunohistochemistry to mealybug squash preparations showed that this protein was a male-specific nuclear protein, but that it was not specifically associated with the heterochromatic set of chromosomes.",
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N2 - The mealybug chromosome cycle is one of the most dramatic examples of genomic imprinting known. In embryos that are to become male the entire paternal chromosome set becomes heterochromatic and inactive at the blastoderm stage, while the maternal set remains active and euchromatic. HP1 is a protein from Drosophila melanogaster, which binds preferentially to heterochromatin on polytene chromosomes and is likely to be a modifier of position effect variegation. This paper describes the isolation and sequencing of two cDNA clones encoding HP1 homologs from the mealybug, Planococcus citri. The protein product of the cDNA clone that was closer to HP1 in sequence was expressed as a fusion protein in Escherichia coli, and polyclonal rat antibodies were raised against it. Immunohistochemistry to mealybug squash preparations showed that this protein was a male-specific nuclear protein, but that it was not specifically associated with the heterochromatic set of chromosomes.

AB - The mealybug chromosome cycle is one of the most dramatic examples of genomic imprinting known. In embryos that are to become male the entire paternal chromosome set becomes heterochromatic and inactive at the blastoderm stage, while the maternal set remains active and euchromatic. HP1 is a protein from Drosophila melanogaster, which binds preferentially to heterochromatin on polytene chromosomes and is likely to be a modifier of position effect variegation. This paper describes the isolation and sequencing of two cDNA clones encoding HP1 homologs from the mealybug, Planococcus citri. The protein product of the cDNA clone that was closer to HP1 in sequence was expressed as a fusion protein in Escherichia coli, and polyclonal rat antibodies were raised against it. Immunohistochemistry to mealybug squash preparations showed that this protein was a male-specific nuclear protein, but that it was not specifically associated with the heterochromatic set of chromosomes.

KW - Amino Acid Sequence

KW - Animals

KW - Base Sequence

KW - Chromosomal Proteins, Non-Histone

KW - Cloning, Molecular

KW - Drosophila melanogaster

KW - Genetic Engineering

KW - Hemiptera

KW - Insect Hormones

KW - Insect Proteins

KW - Molecular Sequence Data

KW - Multigene Family

KW - Nuclear Proteins

KW - Recombinant Fusion Proteins

KW - Sequence Homology, Nucleic Acid

KW - Sex Characteristics

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