Cloning and chromosomal localization of a cDNA encoding a mitochondrial porin from Drosophila melanogaster.
We have raised polyclonal antibodies against purified the Drosphila melanogaster mitochondrial porin. They showed high titre and specificity and were thus used as a tool for screening an expression library. The isolated clone 1T1 showed 74% sequence identity in the last 19 residues at the C-terminus of human porin. A subclone of 1T1, containing the porin-like sequence, was thus used as a probe for re-screening a cDNA library and several positive clones were plaque-purified. We present here the sequence of a 1363 bp cDNA encoding a protein of 279 amino acids. Its identity with porin was also confirmed by N-terminal Edman degradation of the purified protein. The D. melanogaster porin shows an overall 51.8% identity with human porin isoform 1 (porin 31HL or HVDAC1) and an overall 55.7% identity with human porin isoform 2 (HVDAC2). Hydrophobicity plots and secondary structure predictions showed a very high similarity with data obtained from known porin sequences. The D. melanogaster porin cDNA was used as a probe for in situ hybridization to polytenic salivar gland chromosomes. It hybridizes with different intensities in two sites, in chromosome 2L, at region 31E and in chromosome 3L at region 79D. Thus, also in Drosophila melanogaster porin polypeptide(s) belong(s) to a multigene family.
Messina
A
A
Istituto di Scienze Biochimiche e Farmacologiche, Facoltà di Scienze M.F.N., Università di Catania, Italy.
Neri
M
M
Perosa
F
F
Caggese
C
C
Marino
M
M
Caizzi
R
R
De Pinto
V
V
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D016384:Consensus Sequence; D018076:DNA, Complementary; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000235:genetics* / Q000378:metabolism; D015723:Gene Library; D017344:Genes, Insect; D006801:Humans; D007473:Ion Channels / Q000096:biosynthesis / Q000235:genetics; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D008928:Mitochondria / Q000378:metabolism*; D008969:Molecular Sequence Data; D005810:Multigene Family; D009492:Neurospora crassa / Q000235:genetics; D018272:Porins; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid; D011198:Solanum tuberosum / Q000235:genetics; D014908:Triticum / Q000235:genetics; D050995:Voltage-Dependent Anion Channel 1; D050994:Voltage-Dependent Anion Channels
D018076:DNA, Complementary; D029721:Drosophila Proteins; D007473:Ion Channels; D008565:Membrane Proteins; D018272:Porins; D011994:Recombinant Proteins; C494762:VDAC1 protein, human; D050994:Voltage-Dependent Anion Channels; C494761:porin protein, Drosophila; D050995:Voltage-Dependent Anion Channel 1