THE DROSOPHILA PUMILIO GENE ENCODE TWO FUNCTIONAL PROTEIN ISOFORMS THAT PLAY MULTIPLE ROLE IN GERMLINE DEVELOPMENT,

GONADOGENESIS, OOGENESIS AND EMBRYOGENESIS

 The pumilio (pum) play an essential role in

embryonic patterning and germline stem cell maintainance during oogenesis in Drosophila Germline is a unique cell lineage in which a small number of diploid precursor cell undergo drastic proliferation and differentation to produce numerous haploid gametes

 During early oogenesis, pum interact with another

precursor group gene, nanos (nos), to supress the translation of maternally loaded hunchback (hb) transcription factor in the posterior region Pum appears to regulate the proper asymmetric division and maintance the GSC during oogenesis

 Pum protein is present in GSC at higher

conc. Than the cytoblast; various pum mutation affect either the mitotic ability or the depletion of germline during oogenesis Pum encodes at least 2 major funct. protein isoforms 156-kD and a novel 136-kD isoforms

DROSOPHILA STRAIN AND CULTURE

HISTOCHEMISTRY AND IMMUNOFLUORENSCENCE MICROSCOPY

NUCLEIC ACID MANIPULATION

RESCUE OF PUM NUTANT DEFECTS BY PUM CDNA

GERMLINE CLONAL ANALYSIS BY FRT-MEDIATED RECOMBINATION

MRNA IN SITU HYBRIDIZATION

WESTERN BLOTTING ANALYSIS

1) Pum is required for premodial germ cell

development before oogenesis If mutation of pum occur, germline stem cell maintenance during oogenesis will fail Pum ovt mutants Fail to produce any eggs Oogenic defects Ovary size increase Somatis cells number increase

2. Pum 3.

mutants defective in cyst formation and oocyte differentation Defective in oogenesis, but not in ovariole formation Contain few nurse cells but no oocytes because of reduced division of cytoblast. Pum is directly involved in germline stem cell division Mutants produced ovariole that contain only 1-3 mature egg chambers but no other germ cells.

ovt

4. Pum

partially compliment other Pum ovt and maternal effect pum alleles Mutation effects more than maternal effects mutation yet not complementary affect embryonic development. Show maternal effect mutations are stronger mutations Complimentation of Pum 1688 produce viable yet steril progeny

1688

 Pum involve in premordial germ

cell proliferation ovary formation, oogenesis and oviposition Pum does not simply promote or supress the mitotic ability of premodial germ cells Pum required for the growth of premodial germ cell

Pum

is directly required for germline stem cell divisions Pum required for cell maintaining during oogenesis. Pum also required for the differentation or symmetry of germline stem cell division

The 130-kD protein functional

product of pum gene o 130-kD isoform required for germline development & oogenesis in a way similar to that 156-kD o 156-kD and 130-kD proteins possess similar rather than complementary functions

156-kD & 130-kD pum isoforms

posses similar function required for germline development & oogenesis that can be compensated for the dosage-dependant manner Pum continuosly required in germline stem cell division and germline cyst formation during oogenesis

 Either 156-kD or 130-kD

isoforms is sufficient for matenal effect function of pum during embryogenesis

1. The zygotic formation of pum in preoogenic

germline development and oogenesis may require a high dosage of pum expression than the maternal effect function during embryogenesis 2. The maternal loading of both 156-kD and 130kD pum isoforms into the embryo may be abundant so that either isoforms alone can support embrygenesis by providing sufficient dose of pum activity.

The difference between the zygotic and maternal effects may result from the combination of both mechanisms Role of pum in zygotic germline development differs ffrom its maternal effect role in embryonic patterning, where its control the translation of only the hunchback mRNA. I. Pum gene- required for embryonic patterning and germline stem cell division during oogenesis II. Pum func.- primodial germ cell proliferation, ovary formation, oogenesis & ovi-position III. Pum encodes- functional pum isoforms required for developmental processes
3.